Jeffrey S. Church

Environmentally Sustainable Fibers from Regenerated Protein

Concerns for the environment and consumer demand are driving research into environmentally friendly fibers as replacements for part of the 38 million tonnes of synthetic fiber produced annually. While much current research focuses on cellulosic fibers, we highlight that protein fibers regenerated from waste or byproduct sources should also be considered. Feather keratin and wheat gluten may both be suitable. They are annually renewable, commercially abundant, of consistent quality, and have guaranteed supply. They contain useful amino acids for fiber making, with interchain cross-linking possible via cysteine residues or through the metal-catalyzed photocrosslinking of tyrosine residues. Previous commercially produced fibers suffered from poor wet strength. Contemporary nanoparticle and cross-linking technology has the potential to overcome this, allowing commercial production to resume. This would bring together two existing large production and processing pipelines, agricultural protein production and textile processing, to divert potential waste streams into useful products.

Tubular micro-scale multiwalled carbon nanotube-based scaffolds for tissue engineering.

In this study we have prepared a tubular knitted scaffold from a 9 ply multiwalled carbon nanotube (MWCNT) yarn and a composite scaffold, formed by electrospinning poly(lactic-co-glycolic acid) (PLGA) nanofibres onto the knitted scaffold. Both structures were assessed for in vitro biocompatibility with NR6 mouse fibroblast cells for up to 22 days and their suitability as tissue engineering scaffolds considered. The MWCNT yarn was found to support cell growth throughout the culture period, with fibroblasts attaching to, and proliferating on, the yarn surface. The knitted tubular scaffold contained large pores that inhibited cell spanning, leading to the formation of cell clusters on the yarn, and an uneven cell distribution on the scaffold surface. The smaller pores, created through electrospinning, were found to promote cell spanning, leading to a uniform distribution of cells on the composite scaffold surface. Evaluation of the electrical and mechanical properties of the knitted scaffold determined resistance levels of 0.9 kOmega/cm, with a breaking load and extension to break approaching 0.7N and 8%, respectively. The PLGA/MWCNT composite scaffold presented in this work not only supports cell growth, but also has the potential to utilize the full range of electrical and mechanical properties that carbon nanotubes have to offer.

Preparation and characterization of silica coated iron oxide magnetic nano-particles.

Iron oxide magnetic nano-particles have been prepared by precipitation in an aqueous solution of iron(II) and iron(III) chlorides under basic condition. Surface modifications have been carried out by using tetraethoxysilane (TEOS) and mercaptopropyltrimethoxysilane (MPTMS). The uncoated and coated particles have been characterized with transmission electron microscopy (TEM), energy dispersive X-ray (EDX) spectroscopy, thermal gravimetric analysis (TGA), and infrared (IR) and Raman spectroscopy. The particle sizes as measured from TEM images were found to have mean diameters of 13nm for the uncoated and about 19nm for the coated particles. The measured IR spectra of the uncoated and MPTMS coated particles showed the conversion of magnetite to hematite at high temperature. The results obtained from both IR spectroscopy and TGA revealed that the mercaptopropylsilyl group in the MPTMS coated magnetite decomposed at 600 degrees C and the silica layer of the TEOS coated magnetite was rather stable. Raman spectroscopy has shown the laser heating effect through the conversion of magnetite to maghemite and hematite.

The analysis of merino wool cuticle and cortical cells by Fourier transform Raman spectroscopy

Wool fibers are comprised of proteins known as alpha-keratins and have a complex morphological structure. The major components of this structure, the cuticle and cortical cells, differ in the conformations of their chains as well as their amino acid compositions. High quality Fourier transform Raman spectra of cortical and cuticle cells isolated from fine Merino wool fibers have been obtained. Raman spectroscopy has been shown to be sensitive to the differences in both secondary structure and amino acid composition. The cortical cells were found to be higher in alpha-helical content as compared to the cuticle cells, which had an increased disordered content. Specific information, consistent with amino acid analysis results, regarding cystine, tyrosine, tryptophan, and phenylalnine residues, were obtained for both the cortical and cuticle cells. In addition, the Raman spectra provided information about free thiol groups, amino acids residues with amide group side chains, and residues with protonated carboxyl group side chains. Middle ir transmission spectra of these isolated cells were also obtained. In comparison to the Raman data, the middle ir spectra were found to be not as rich in information.

Raman spectroscopy in the analysis of food and pharmaceutical nanomaterials

Raman scattering is an inelastic phenomenon. Although its cross section is very small, recent advances in electronics, lasers, optics, and nanotechnology have made Raman spectroscopy suitable in many areas of application. The present article reviews the applications of Raman spectroscopy in food and drug analysis and inspection, including those associated with nanomaterials. Brief overviews of basic Raman scattering theory, instrumentation, and statistical data analysis are also given. With the advent of Raman enhancement mechanisms and the progress being made in metal nanomaterials and nanoscale metal surfaces fabrications, surface enhanced Raman scattering spectroscopy has become an extra sensitive method, which is applicable not only for analysis of foods and drugs, but also for intracellular and intercellular imaging. A Raman spectrometer coupled with a fiber optics probe has great potential in applications such as monitoring and quality control in industrial food processing, food safety in agricultural plant production, and convenient inspection of pharmaceutical products, even through different types of packing. A challenge for the routine application of surface enhanced Raman scattering for quantitative analysis is reproducibility. Success in this area can be approached with each or a combination of the following methods: (1) fabrication of nanostructurally regular and uniform substrates; (2) application of statistic data analysis; and (3) isotopic dilution.

Honeybee silk: Recombinant protein production, assembly and fiber spinning

Transgenic production of silkworm and spider silks as biomaterials has posed intrinsic problems due to the large size and repetitive nature of the silk proteins. In contrast the silk of honeybees (Apis mellifera) is composed of a family of four small and non-repetitive fibrous proteins. We report recombinant production and purification of the four full-length unmodified honeybee silk proteins in Escherichia coli at substantial yields of 0.2-2.5 g/L. Under the correct conditions the recombinant proteins self-assembled to reproduce the native coiled coil structure. Using a simple biomimetic spinning system we could fabricate recombinant silk fibers that replicated the tensile strength of the native material.

Characterization of oxides on niobium by raman and infrared spectroscopy

The combination of electrochemical methods, laser Raman spectroscopy (LRS), surface-enhanced Raman scattering (SERS) and infrared reflection absorption spectroscopy (IRRAS) have been used to demonstrate that a stable passivated film can easily be formed on Nb in 0.15 M NaCl. The composition of the film has been identified to be NbO2 and different forms of Nb2O5. From LRS, the film formed in 0.10 M NaOH has similar composition. A longer exposure of the Nb electrode to air favors the conversion of NbO2 to the more stable Nb2O5 and the transformation of BNb2O5 to HNb2O5. In situ analysis of the passivated film in 0.15 M NaCl was carried out by coupling the electrochemical technique with LRS and SERS. The results indicated the presence of different forms of Nb2O5 at the metal/solution interfaces.

Single Honeybee Silk Protein Mimics Properties of Multi-Protein Silk

Honeybee silk is composed of four fibrous proteins that, unlike other silks, are readily synthesized at full-length and high yield. The four silk genes have been conserved for over 150 million years in all investigated bee, ant and hornet species, implying a distinct functional role for each protein. However, the amino acid composition and molecular architecture of the proteins are similar, suggesting functional redundancy. In this study we compare materials generated from a single honeybee silk protein to materials containing all four recombinant proteins or to natural honeybee silk. We analyse solution conformation by dynamic light scattering and circular dichroism, solid state structure by Fourier Transform Infrared spectroscopy and Raman spectroscopy, and fiber tensile properties by stress-strain analysis. The results demonstrate that fibers artificially generated from a single recombinant silk protein can reproduce the structural and mechanical properties of the natural silk. The importance of the four protein complex found in natural silk may lie in biological silk storage or hierarchical self-assembly. The finding that the functional properties of the mature material can be achieved with a single protein greatly simplifies the route to production for artificial honeybee silk.

FT-Raman spectroscopy of wool—I. Preliminary studies

Abstract The FT-Raman spectrum of wool has been obtained using near-IR excitation. No significant fluorescence was observed and the spectra could be obtained routinely. No sample damage was observed for laser powers up to 400 mW. Several methods of sample presentation for both wet and dry wool were investigated and the optimum data collection conditions were determined for each. Dry dense plugs of wool fibers (∼0.16 g cm −3 ) were found to provide the best spectra. Due to improvements in band resolution and signal-to-noise ratio, several previously unobserved spectral features in the wool spectrum have become apparent. The assignment of the Raman-active vibrational modes of wool are reviewed and updated to include these features. Raman spectra obtained from chlorinated and untreated wool samples did not exhibit any significant differences. In contrast, the ATR spectra obtained from these samples exhibited significant differences in the SO stretching region.

An Unlikely Silk: The Composite Material of Green Lacewing Cocoons

Spiders routinely produce multiple types of silk; however, common wisdom has held that insect species produce one type of silk each. This work reports that the green lacewing ( Mallada signata, Neuroptera) produces two distinct classes of silk. We identified and sequenced the gene that encodes the major protein component of the larval lacewing cocoon silk and demonstrated that it is unrelated to the adult lacewing egg-stalk silk. The cocoon silk protein is 49 kDa in size and is alanine rich (>40%), and it contains an alpha-helical secondary structure. The final instar lacewing larvae spin protein fibers of approximately 2 microm diameter to construct a loosely woven cocoon. In a second stage of cocoon construction, the insects lay down an inner wall of lipids that uses the fibers as a scaffold. We propose that the silk protein fibers provide the mechanical strength of the composite lacewing cocoon whereas the lipid layer provides a barrier to water loss during pupation.