Melt electrospinning writing

This thesis has developed an innovative additive manufacturing technology platform, which combines melt electrospinning with direct writing, allowing the fabrication of a new class of highly-ordered ultrafine fibrous materials. Bioresorbable polymer fibres were printed using a variety of designs, with filament resolutions not demonstrated by established melt-extrusion based direct writing processes, to form novel medical devices. This platform was applied to tissue engineering scaffold design, where structures were prepared in a variety of shapes and forms, characterised and then seeded with cells to investigate their biocompatibility, cell-seeding and proliferation behaviour as well as the ability to guide cell growth and differentiation.

Fabrication using a rapid prototyping system and in vitro characterization of PEG-PCL-PLA scaffolds for tissue engineering

n the field of tissue engineering new polymers are needed to fabricate scaffolds with specific properties depending on the targeted tissue. This work aimed at designing and developing a 3D scaffold with variable mechanical strength, fully interconnected porous network, controllable hydrophilicity and degradability. For this, a desktop-robot-based melt-extrusion rapid prototyping technique was applied to a novel tri-block co-polymer, namely poly(ethylene glycol)-block-poly(epsi-caprolactone)-block-poly(DL-lactide), PEG-PCL-P(DL)LA. This co-polymer was melted by electrical heating and directly extruded out using computer-controlled rapid prototyping by means of compressed purified air to build porous scaffolds. Various lay-down patterns (0/30/60/90/120/150°, 0/45/90/135°, 0/60/120° and 0/90°) were produced by using appropriate positioning of the robotic control system. Scanning electron microscopy and micro-computed tomography were used to show that 3D scaffold architectures were honeycomb-like with completely interconnected and controlled channel characteristics. Compression tests were performed and the data obtained agreed well with the typical behavior of a porous material undergoing deformation. Preliminary cell response to the as-fabricated scaffolds has been studied with primary human fibroblasts. The results demonstrated the suitability of the process and the cell biocompatibility of the polymer, two important properties among the many required for effective clinical use and efficient tissue-engineering scaffolding.

Polycaprolactone-based scaffold plus rhBMP-2 in a sheep thoracic spine fusion model

We report the application of a novel scaffold design in a sheep thoracic spine model for spine deformity correction. The combination of the calcium-phosphate coated polycaprolactone scaffolds with recombinant human bone morphogenic protein-2 (rhBMP-2) are intended as a future bone graft substitute in ensuring the stability of bony intervertebral fusion. A solid free-form fabrication process based on melt extrusion has been utilized in the manufacturing of these scaffolds. To date there are no studies examining the use of such biodegradable implants in a sheep thoracic spine model. The success of anterior scoliosis surgery in humans depends on achieving a solid bony fusion between adjacent vertebrae after the intervertebral discs have been surgically cleared and the disc spaces filled with graft material. Due to limited availability of autograft, there is much current interest in the development of synthetic scaffolds in combination with growth factors such as recombinant human bone morphogenetic protein (rhBMP-2) to achieve a solid bony fusion following scoliosis surgery.

Environmental degradation of lignin/poly(hydroxybutyrate) blends

Blends of lignin and poly(hydroxybutyrate) (PHB) were obtained by melt extrusion. They were buried in a garden soil for up to 12 months, and the extent and mechanism of degradation were investigated by gravimetric analysis, thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), X-ray photoelectron spectroscopy (XPS), scanning electron microscopy (SEM) and Fourier transform infra-red spectroscopy (FTIR) over the entire range of compositions. The PHB films were disintegrated and lost 45 wt% of mass within 12 months. This value dropped to 12 wt% of mass when only 10 wt% of lignin was present, suggesting that lignin both inhibited and slowed down the rate of PHB degradation. TGA and DSC indicated structural changes, within the lignin/PHB matrix, with burial time, while FTIR results confirmed the fragmentation of the PHB polymer. XPS revealed an accumulation of biofilms on the surface of buried samples, providing evidence of a biodegradation mechanism. Significant surface roughness was observed with PHB films due to microbial attack caused by both loosely and strongly associated micro-organisms. The presence of lignin in the blends may have inhibited the colonisation of the micro-organisms and caused the blends to be more resistant to microbial attack. Analysis suggested that lignin formed strong hydrogen bonds with PHB in the buried samples and it is likely that the rate of breakdown of PHB is reduced, preventing rapid degradation of the blends.

Design and characterisation of scaffolds for osteochondral regeneration

Treatment of joint diseases such as osteoarthritis is difficult and requires extensive developments for adequate solutions to emerge. Continued innovation in projects explored in this thesis may be beneficial to understanding the requirements of the joint environment. This may then lead to constructs that perform desirably from both mechanical and biological standpoints, resulting in complete, tissue-engineered osteochondral solutions. This thesis investigated specific scaffold designs for bone and osteochondral tissue engineering, as well as the formation of complex criteria on which cartilage hydrogel scaffolds may be assessed. The combination of hydrogels and ceramics were found to maintain chondrogenesis, while the concentration of photoinitiators in photocrosslinkable hydrogel systems may be optimised to maximise mechanical properties and cell viability. Finally, viscoelasticity of hydrogel blends was assessed using oscillatory motion, demonstrating the property is tailorable.

Vibrational spectroscopic studies of laboratory scale polymer melt processing : application to a thermoplastic polyurethane nanocomposite

A laboratory scale twin screw extruder has been interfaced with a near infrared (NIR) spectrometer via a fibre optic link so that NIR spectra can be collected continuously during the small scale experimental melt state processing of polymeric materials. This system can be used to investigate melt state processes such as reactive extrusion, in real time, in order to explore the kinetics and mechanism of the reaction. A further advantage of the system is that it has the capability to measure apparent viscosity simultaneously which gives important additional information about molecular weight changes and polymer degradation during processing. The system was used to study the melt processing of a nanocomposite consisting of a thermoplastic polyurethane and an organically modified layered silicate.

Melt electrospinning of polycaprolactone and its blends with poly(ethylene glycol)

Melt electrospinning is one aspect of electrospinning with relatively little published literature, although the technique avoids solvent accumulation and/or toxicity which is favoured in certain applications. In the study reported, we melt-electrospun blends of poly(ε-caprolactone) (PCL) and an amphiphilic diblock copolymer consisting of poly(ethylene glycol) and PCL segments (PEG-block-PCL). A custom-made electrospinning apparatus was built and various combinations of instrument parameters such as voltage and polymer feeding rate were investigated. Pure PEG-block-PCL copolymer melt electrospinning did not result in consistent and uniform fibres due to the low molecular weight, while blends of PCL and PEG-block-PCL, for some parameter combinations and certain weight ratios of the two components, were able to produce continuous fibres significantly thinner (average diameter of ca 2 µm) compared to pure PCL. The PCL fibres obtained had average diameters ranging from 6 to 33 µm and meshes were uniform for the lowest voltage employed while mesh uniformity decreased when the voltage was increased. This approach shows that PCL and blends of PEG-block-PCL and PCL can be readily processed by melt electrospinning to obtain fibrous meshes with varied average diameters and morphologies that are of interest for tissue engineering purposes. Copyright © 2010 Society of Chemical Industry

Melt electrospinning

Melt electrospinning is relatively under-investigated compared to solution electrospinning but provides opportunities in numerous areas, in which solvent accumulation or toxicity are a concern. These applications are diverse, and provide a broad set of challenges to researchers involved in electrospinning. In this context, melt electrospinning provides an alternative approach that bypasses some challenges to solution electronspinning, while bringing new issues to the forefront, such as the thermal stability of polymers. This Focus Review describes the literature on melt electrospinning, as well as highlighting areas where both melt and solution are combined, and potentially merge together in the future.

High-resolution DNA melt curve analysis of the clustered, regularly interspaced short-palindromic-repeat locus of Campylobacter jejuni

A novel method for genotyping the clustered, regularly interspaced short-palindromic-repeat (CRISPR) locus of Campylobacter jejuni is described. Following real-time PCR, CRISPR products were subjected to high-resolution melt (HRM) analysis, a new technology that allows precise melt profile determination of amplicons. This investigation shows that the CRISPR HRM assay provides a powerful addition to existing C. jejuni genotyping methods and emphasizes the potential of HRM for genotyping short sequence repeats in other species

Preliminary validation of a novel high-resolution melt-based typing method based on the multilocus sequence typing scheme of Streptococcus pyogenes

The major limitation of current typing methods for Streptococcus pyogenes, such as emm sequence typing and T typing, is that these are based on regions subject to considerable selective pressure. Multilocus sequence typing (MLST) is a better indicator of the genetic backbone of a strain but is not widely used due to high costs. The objective of this study was to develop a robust and cost-effective alternative to S. pyogenes MLST. A 10-member single nucleotide polymorphism (SNP) set that provides a Simpson’s Index of Diversity (D) of 0.99 with respect to the S. pyogenes MLST database was derived. A typing format involving high-resolution melting (HRM) analysis of small fragments nucleated by each of the resolution-optimized SNPs was developed. The fragments were 59–119 bp in size and, based on differences in G+C content, were predicted to generate three to six resolvable HRM curves. The combination of curves across each of the 10 fragments can be used to generate a melt type (MelT) for each sequence type (ST). The 525 STs currently in the S. pyogenes MLST database are predicted to resolve into 298 distinct MelTs and the method is calculated to provide a D of 0.996 against the MLST database. The MelTs are concordant with the S. pyogenes population structure. To validate the method we examined clinical isolates of S. pyogenes of 70 STs. Curves were generated as predicted by G+C content discriminating the 70 STs into 65 distinct MelTs.

Design and fabrication of tubular scaffolds via direct writing in a melt electrospinning mode

Flexible tubular structures fabricated from solution electrospun fibers are finding increasing use in tissue engineering applications. However it is difficult to control the deposition of fibers due to the chaotic nature of the solution electrospinning jet. By using non-conductive polymer melts instead of polymer solutions the path and collection of the fiber becomes predictable. In this work we demonstrate the melt electrospinning of polycaprolactone in a direct writing mode onto a rotating cylinder. This allows the design and fabrication of tubes using 20 μm diameter fibers with controllable micropatterns and mechanical properties. A key design parameter is the fiber winding angle, where it allows control over scaffold pore morphology (e.g. size, shape, number and porosity). Furthermore, the establishment of a finite element model as a predictive design tool is validated against mechanical testing results of melt electrospun tubes to show that a lesser winding angle provides improved mechanical response to uniaxial tension and compression. In addition, we show that melt electrospun tubes support the growth of three different cell types in vitro and are therefore promising scaffolds for tissue engineering applications.

Melt textured Y123 bulk and thick film

YBa2Cu3O7-δ - 25mol%Y2BaCuO5 bars and thick films have been melt textured using a stationary furnace with a temperature gradient of 3 or 6°C/cm. Samples are heated above the peritectic reaction temperature and quenched to just above the solidification temperature and then slowly cooled below the solidification temperature. All bar shaped samples consist of 2-5 mm grains though the grain orientations strongly depend on the heat treatment conditions. The bar shows the maximum Jc of above 3,000 A/cm2, whereas the maximum Jc of 200 A/cm2 and Tczero of 88K are obtained for the thick film on (100) LaAlO3 single crystal.

Melt-textured YBa2Cu3O7-x wires having plate-like grains

YBCO wires which consist of well oriented plate-like fine grains are fabricated using a moving furnace to achieve higher mechanical strength. Melt-texturing experiments have been undertaken on YBCO wires with two different compositions: YBa1.5Cu2.9O7-x, and YBa1.8Cu3.0O7-x. Wires are extruded from a mixture of precursor powders (formed by a coprecipitation process) then textured by firing in a moving furnace. Size of secondary phases such as barium cuprate and copper oxide, and overall composition of the sample affect the orientation of the fine grains. At zero magnetic field, the YBa1.5Cu2.9O7-x wire shows the highest critical current density of 1,450 Acm-2 and 8,770 Acm-2 at 77K and 4.2K, respectively. At 1 T, critical current densities of 30 Acm-2 and 200 Acm-2, respectively, are obtained at 77K and 4.2K. Magnetisation curves are also obtained for one sample to evaluate critical current density using the Bean model. Analysis of the microstructure indicates that the starting composition of the green body significantly affects the achievement of grain alignment via melt-texturing processes.