Thèses sur le sujet « Melt electrospinning »

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.

In this thesis, the suitability of Melt Electrospinning Writing technology is demonstrated for two applications: building functional tissue substitutes and engineering relevant models to study disease mechanisms. More specifically, a myocardial patch was built for cardiac tissue engineering, and a prostate microtissue was engineered to study the interactions between epithelium and stroma during prostate cancer progression. This thesis corresponds to the dual Master in Biofabrication, carried out between Universiteit Utrecht and Queensland University of Technology.

This thesis was a study into the effect of charge buildup and subsequent modifications on a direct writing melt electrospinning device. It examined the effect of distributing the application of charge in a melt electrospinning system and studying the effect this had on order in scaffolds produced. The distribution of charge led to an increase in deposition accuracy in layers 10 times higher than previously achievable.

This project was a step forward in developing an image-based control environment for an additive manufacturing process using polymer melts. The monitoring system provides a real time footage, that allows for analysis of the process and control of the morphology of printed structures. The system was used to conduct studies focused on the effect of printing conditions on the fibre's morphology and behaviour as well as provides the basis for future development towards quality control and optimisation.

The addition of three-dimensional structure in hydrogels, and the first reported instance of melt electrospinning writing (MEW) of polypropylene, provides a foundation for the production of complex hydrogel systems for a variety of applications. This project provides a novel, facile and universal method to produce porous structures in soft hydrogels, using sacrificial templates produced via MEW. While the optimization of the MEW of polypropylene was completed elucidating methods to enable the production of MEW scaffolds with high melting point polymers.

This thesis developed a method for designing 3D printed implants to restore bone loss. Using melt-electrospinning 3D printing technology and patient medical scan data, the researcher designed and fabricated anatomically-accurate scaffolds using biodegradable polymers to ultimately facilitate bone regeneration. The method presented was applied to three clinically-relevant case studies and can now be used for the design of a range of other implants based on patient scan data. The application and importance of this method was discussed as a key element in the biofabrication process for the fabrication of biologically-relevant, patient-specific human tissues and organs.

This thesis was a step forward in the development of an effective and patient-specific treatment for bone tissue loss using synthetic tissue engineered constructs. A novel polycaprolactone/strontium-substituted bioactive glass composite was fabricated into scaffolds with highly ordered fibre structure and promising osteogenic potential using an advanced additive manufacturing technique known as melt-electrospinning. The findings of this thesis provide an indispensable link in our understanding of future cell-free treatment for bone defects utilising fully synthetic bioactive scaffolds. The thesis also developed several histological assessment tools for evaluating current and future tissue engineered bone constructs utilised in pre-clinical animal studies.

This thesis establishes an additive biomanufacturing technology platform for melt electrowriting. It has been hypothesised that applying systems engineering methodologies assists in developing control, reproducibility, and scale. The implementation of automated monitoring and parameter control helped to generate large data to identify the optimum settings. A conclusion is drawn between the geometry of a fibre and the quality of morphology, assessing robustness and reproducibility. Additionally, contemporary achievable scaffold fabrication heights of 2 mm to 7 mm is achieved and the influence of gravity is evaluated. The resulting technologies are utilised to design and develop a high-throughput printer.

The thesis deals with the principal heating design of the electrospinning electrode. These electrodes are fed with high voltage (tens of kV), which is an essential part of the device for the nanofibres production. This thesis presents the research of possible solutions of the heating and also it analyzes some principles of the nanofibre production. Furthermore, the work presents electrical schematics and PCB of the heating module. The aim of this work is to propose such a solution of heating that makes spinning from melt realizable in the device for 4SPIN ® nanofibres production.

Design strategies inspired by nature open up new avenues in materials design and facilitate the development of innovative materials outperforming their conventionally engineered counterparts. In this thesis, bioinspired design principles based on the physicochemical and morphological properties of soft biological materials were used to develop functional soft network composites (SNCs) intended for soft tissue engineering applications. These SNCs consist of a network of 3D printed microfibres and a hydrogel matrix mimicking the collagens and proteoglycans present in native extracellular matrices, respectively. Our results suggest that this new class of composites are suitable for tissue engineering a broad range of soft tissues including cartilage, skin, ligament, tendon, muscle and heart valve.

In order to improve the purity of steel castings, the use of special reactive coatings on carbon-bonded ceramic foam filters was explored. Carbon nanotubes were dispersed in water by means of ultrasonic treatment, using xanthan gum to stabilize the nanotubes in suspension and control the rheological behavior. The coatings were applied by cold spraying and binding was achieved during heat treatment in reducing atmosphere, thanks to an artificial pitch added to the slurry. The coated filters were successfully immersed in molten steel for different times. The thickness of the first alumina layer generated at the interface was independent of the immersion time: concentration gradients through its thickness suggested that the formation of this structure is limited by diffusion. Investigation of the steel after solidification by means of ASPEX showed that the presence of the coating influenced the size as well as the chemical composition of the remaining inclusions. Nano-coated filters had the best filtration efficiency (up to 95% for alumina inclusions after 10 s), but longer tests resulted in worse performance. In addition, coatings based on calcium aluminates in combination with carbon showed an efficiency greater than 97% for steel samples taken directly from the melt.

This thesis focuses on the development of a humanised mouse model to investigate human breast cancer metastasis to bone, an incurable disease presenting a major medical challenge in our society. The method is based on tissue-engineered constructs with human cells that generate a human bone-like organ within mice. This novel platform is further applied to mimic human-specific mechanisms of breast cancer metastasis and growth in human bone, and in particular the role of specific cell adhesion molecules in this process is closely investigated.

This research was focused on the development of advanced quantitative methods to evaluate the use of a 3D printed membrane scaffold, for the guidance and spatiotemporal delivery of recombinant human bone growth factor for the regeneration of a large bone defect, which still represents a major challenge in orthopaedic and reconstructive surgery. Two histomorphometric approaches were investigated and successfully applied. The combined histological, immunohistochemical and histomorphometric analyses outlined in this thesis further elucidated the mechanisms and patterns that govern the bone regeneration of a critical sized bone defect in a large experimental ovine model.

Synthetic hydrogels featuring tunable biological functionalities and hierarchical structures are of compelling interest as scaffolds for tissue engineering applications. With the expectation of regulating cell fate within the soft materials, many efforts have been placed on creating niches that can mimic the complexity of the native extracellular matrix. In this study, a sacrificial moulding process was used to produce porous hydrogels, while two patterning approaches were developed to site-specifically immobilize molecules inside the hydrogels, resembling natural extracellular matrix networks in terms of geometrical interconnectivity and cell-guidance functionalization. The simple approaches allow reproducible control over the size and architecture of the channels, as well as the spatial distribution and concentration of the patterning molecules, enabling controlled study of cell-substrate behaviour.

This thesis comprises two research projects undertaken as part of the dual biofabrication master's program between Queensland University of Technology and Utrecht University. The two projects focused on leveraging biofabrication, tissue-culture, and soft robotics to develop novel methods for fabricating 3D vascular and colon tissue, respectively. The first project developed a novel approach for conditioning cells using soft robotics that emulate vascular biomechanics, whereas the second project combined bioink micromoulding and melt electrospinning writing to fabricate 3D colon organoid constructs that mimic colon crypt morphology. Together, these projects contribute innovative biofabrication methods for creating tissue-culture models with enhanced biomimicry.

This project designed and developed a novel in vitro model of the haematopoietic stem cell niche. Components of the endosteal niche and the perivascular niche, essential in the bone marrow haematopoietic stem cell niche microenvironment, were integrated in a single platform using a multiphasic approach that combined melt electrospun written scaffolds with starPEG-heparin hydrogels. Haematopoietic stem cell response was analysed after 3D co-culture with the tissue engineering niches.

Tissue engineering provides a potential solution for the repair and regeneration of bone defects and fractures healing. A biomedical scaffold is one of the ideal approaches to achieve effective structure for bone cell growth and bone formation in the desired shape. This study has developed an ideal three-dimensional scaffold architecture with improved biological functionality, which has a physically stable and structurally porous shape, with interconnected channels and defined topography for guided bone regeneration.

El presente trabajo tiene por objetivo estudiar las aplicaciones de los nanocristales o ¿nanowhiskers¿ extraídos mediante hidrólisis ácida de celulosa bacteriana (BCNW) para el desarrollo de materiales poliméricos y biopoliméricos con propiedades mejoradas para su uso en aplicaciones de envasado de alimentos. En primer lugar se estudió y optimizó el proceso de extracción de BCNW. Se desarrolló un procedimiento de extracción con ácido sulfúrico, que permitió obtener nanocristales con elevada relación de aspecto y cristalinidad y al mismo tiempo, un elevado rendimiento de extracción. Este procedimiento comprende una posterior etapa de neutralización que resultó ser necesaria para garantizar la estabilidad térmica de los nanocristales. El siguiente paso consistió en la formulación de materiales nanocompuestos con propiedades mejoradas incorporando BCNW en diferentes matrices plásticas, en concreto copolímeros de etileno-alcohol vinílico (EVOH), ácido poliláctico (PLA) y polihidroxialcanoatos (PHAs). Mediante las técnicas de electroestirado y estirado por soplado se generaron fibras híbridas de EVOH y PLA con BCNW. La incorporación de BCNW en las disoluciones empleadas para producir fibras modificó significativamente sus propiedades (viscosidad, tensión superficial y conductividad) y por tanto, la morfología de las fibras se vio afectada. Además, se generaron fibras con propiedades antimicrobianas mediante la incorporación de aditivos, maximizando el efecto antimicrobiano con la adición de sustancias de carácter hidrofílico. Seguidamente, se produjeron nanocompuestos por mezclado en fundido y se desarrollaron técnicas de pre-incorporación de BCNW para evitar la aglomeración de los mismos no sólo en matrices hidrofílicas como el EVOH, sino también en matrices hidrofóbicas como el PLA. La dispersión óptima de BCNW resultó en una mejora de las propiedades mecánicas y de barrera de los nanocompuestos. También se estudió la modificación de la superficie de los nanocristales mediante copolimerización con poli(glicidil metacrilato) para mejorar la compatibilidad de BCNW con una matriz hidrofóbica como el PLA. Se incluyen además los primeros resultados obtenidos en cuanto a la producción de nanobiocompuestos sintetizados por microorganismos, que consisten en PHAs con diferentes contenidos de hidroxivalerato reforzados con BCNW. Por último, se desarrollaron películas con propiedades de alta barrera basadas en películas de BCNW recubiertas con capas hidrofóbicas. El recubrimiento mediante la deposición de fibras por electrospinning seguido de homogeneización por calentamiento garantizó una buena adhesión entre las diferentes capas, protegiendo así las películas de BCNW del efecto negativo de la humedad.
Martínez Sanz, M. (2013). Bacterial cellulose nanowhiskers to enhance the properties of plastics and bioplastics of interest in food packaging [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/30312
TESIS
Premiado

In order to mimic the extracellular matrix for tissue engineering, recent research approaches often involve 3D printing or electrospinning of fibres to scaffolds as cell carrier material. Within this thesis, a micron fibre printing process, called melt electrospinning writing (MEW), combining both additive manufacturing and electrospinning, has been investigated and improved. Thus, a unique device was developed for accurate process control and manufacturing of high quality constructs. Thereby, different studies could be conducted in order to understand the electrohydrodynamic printing behaviour of different medically relevant thermoplastics as well as to characterise the influence of MEW on the resulting scaffold performance. For reproducible scaffold printing, a commonly occurring processing instability was investigated and defined as pulsing, or in extreme cases as long beading. Here, processing analysis could be performed with the aim to overcome those instabilities and prevent the resulting manufacturing issues. Two different biocompatible polymers were utilised for this study: poly(ε-caprolactone) (PCL) as the only material available for MEW until then and poly(2-ethyl-2-oxazoline) for the first time. A hypothesis including the dependency of pulsing regarding involved mass flows regulated by the feeding pressure and the electrical field strength could be presented. Further, a guide via fibre diameter quantification was established to assess and accomplish high quality printing of scaffolds for subsequent research tasks. By following a combined approach including small sized spinnerets, small flow rates and high field strengths, PCL fibres with submicron-sized fibre diameters (fØ = 817 ± 165 nm) were deposited to defined scaffolds. The resulting material characteristics could be investigated regarding molecular orientation and morphological aspects. Thereby, an alignment and isotropic crystallinity was observed that can be attributed to the distinct acceleration of the solidifying jet in the electrical field and by the collector uptake. Resulting submicron fibres formed accurate but mechanically sensitive structures requiring further preparation for a suitable use in cell biology. To overcome this handling issue, a coating procedure, by using hydrophilic and cross-linkable star-shaped molecules for preparing fibre adhesive but cell repellent collector surfaces, was used. Printing PCL fibre patterns below the critical translation speed (CTS) revealed the opportunity to manufacture sinusoidal shaped fibres analogously to those observed using purely viscous fluids falling on a moving belt. No significant influence of the high voltage field during MEW processing could be observed on the buckling phenomenon. A study on the sinusoidal geometry revealed increasing peak-to-peak values and decreasing wavelengths as a function of decreasing collector speeds sc between CTS > sc ≥ 2/3 CTS independent of feeding pressures. Resulting scaffolds printed at 100 %, 90 %, 80 % and 70 % of CTS exhibited significantly different tensile properties, foremost regarding Young’s moduli (E = 42 ± 7 MPa to 173 ± 22 MPa at 1 – 3 % strain). As known from literature, a changed morphology and mechanical environment can impact cell performance substantially leading to a new opportunity of tailoring TE scaffolds. Further, poly(L-lactide-co-ε-caprolactone-co-acryloyl carbonate) as well as poly(ε-caprolactone-co-acryloyl carbonate) (PCLAC) copolymers could be used for MEW printing. Those exhibit the opportunity for UV-initiated radical cross-linking in a post-processing step leading to significantly increased mechanical characteristics. Here, single fibres of the polymer composed of 90 mol.% CL and 10 mol.% AC showed a considerable maximum tensile strength of σmax = 53 ± 16 MPa. Furthermore, sinusoidal meanders made of PCLAC yielded a specific tensile stress-strain characteristic mimicking the qualitative behaviour of tendons or ligaments. Cell viability by L929 murine fibroblasts and live/dead staining with human mesenchymal stem cells revealed a promising biomaterial behaviour pointing out MEW printed PCLAC scaffolds as promising choice for medical repair of load-bearing soft tissue. Indeed, one apparent drawback, the small throughput similar to other AM methods, may still prevent MEW’s industrial application yet. However, ongoing research focusses on enlargement of manufacturing speed with the clear perspective of relevant improvement. Thereby, the utilisation of large spinneret sizes may enable printing of high volume rates, while downsizing the resulting fibre diameter via electrical field and mechanical stretching by the collector uptake. Using this approach, limitations of FDM by small nozzle sizes could be overcome. Thinking visionary, such printing devices could be placed in hospitals for patient-specific printing-on-demand therapies one day. Taking the evolved high deposition precision combined with the unique small fibre diameter sizes into account, technical processing of high performance membranes, filters or functional surface finishes also stands to reason
Um biomimetische extrazelluläre Matrices für das Tissue Engineering herzustellen, bedienen sich aktuelle Forschungsansätze oftmals der Produktion von Faser-Konstrukten durch additive Fertigung oder Elektrospinn-Verfahren. Das sogenannte Melt Electrospinning Writing (MEW) kombiniert Vorteile beider Techniken und weist dadurch ein hohes Applikationspotential auf. Daher bestand das Ziel der vorliegenden Arbeit in der Weiterentwicklung und Erforschung des MEW. Für diesen Zweck wurde eine neuartige Forschungsanlage konzipiert und gebaut, welche mit einzigartiger Verfahrenspräzision und Prozesskontrolle die Fertigung von hochqualitativen Konstrukten ermöglichte. Auf Basis dessen konnten die durchgeführten Studien das Verständnis des elektrohydrodynamischen Druckvorgangs und der untersuchten Prozessparameter vertiefen und letztendlich zur Ausweitung des Verfahrens auf neue medizinisch relevante Thermoplaste beitragen. Um eine reproduzierbare Herstellung von Scaffolds zu ermöglichen, wurde eine häufig auftretende Prozessinstabilität erforscht und als pulsing, oder in stark ausgeprägten Fällen als long beading, klassifiziert. Durch Prozessanalyse konnte zudem eine Methode zur Vermeidung dieser Instabilität entwickelt werden. Dafür wurden zwei unterschiedliche biokompatible Polymere verwendet: Poly(ε-Caprolacton) (PCL) als bis dahin einziger verfügbarer MEW Werkstoff, sowie erstmalig Poly(2-Ethyl-2-Oxazolin). Die aufgestellte Hypothese umfasst eine universelle Abhängigkeit der pulsing Instabilität zu involvierten Massenströmen, welche durch Anpassung des angelegten Prozessdruckes und der elektrischen Feldstärke reguliert werden kann. Um ein optimales Prozessergebnis für nachfolgende Forschungsarbeiten zu erzielen, wurde zusätzlich ein Leitfaden zur quantitativen Bewertung des Grades der Instabilität bereitgestellt. Durch Kombination kleiner Spinndüsen, kleiner Schmelze-Flussraten und hoher elektrischen Feldstärken, konnten erstmalig PCL Fasern mit sub-mikron Durchmessern (fØ = 817 ± 165 nm) zu präzisen Scaffolds verarbeitet werden. Diese wurden anschließend durch materialwissenschaftliche Analytik charakterisiert. Dabei wurde eine molekulare Vorzugsorientierung und isotrope Kristallausrichtung entlang der Faser beobachtet, welche durch den hohen Verstreckungsgrad des erstarrenden Polymerstrahls erklärt werden konnte. Resultierende sub-mikron Fasern konnten zwar für einen akkuraten Druckvorgang verwendet werden, jedoch erwiesen sich die Strukturen als instabil und daher nicht geeignet für die Handhabung bei Zellkulturstudien. Aus diesem Grund wurde ein Beschichtungsansatz mittels hydrophilen und vernetzbaren Sternmolekülen für Substratflächen herangezogen. Während solche modifizierten Oberflächen bekanntermaßen Zelladhäsion verhindern, konnten gedruckte sub-mikron Scaffolds auf der Oberfläche haften und so für biologische Studien verwendet werden. Durch das gezielte Ablegen von Fasern unterhalb der kritischen Translationsgeschwindigkeit (CTS) des Kollektors, konnten sinusförmige Faserstrukturen erzeugt werden. Analog zu rein viskosen Fluiden, welche durch ein bewegliches Band aufgesammelt werden, schien dieser Vorgang dem sogenannten buckling zu unterliegen und daher phänomenologisch nicht oder nur geringfügig vom elektrischen Feld abhängig zu sein. Zudem konnte eine durchgeführte Studie die direkte Abhängigkeit der Fasergeometrie mit der Kollektorbewegung belegen. Unabhängig vom Prozessdruck, führte eine verminderte Kollektorgeschwindigkeit sc in den Grenzen CTS > sc ≥ 2/3 CTS zu erhöhten Amplituden bzw. Spitze-zu-Spitze Werten und verkürzten Wellenlängen. Durch das kontrollierte Ablegen der Fasern bei Geschwindigkeiten von 100 %, 90 % 80 % und 70 % CTS konnten zudem Scaffolds mit unterschiedlichen mechanischen Eigenschaften hergestellt werden. Speziell der Zugmodul wurde dadurch etwa um eine halbe Größenordnung moduliert (Es = 42 ± 7 MPa bis 173 ± 22 MPa bei 1 – 3 % Dehnung). Dies ist in Kombination mit der Strukturierung für maßgeschneiderte TE Scaffolds von großem Interesse, da zelluläre Systeme sensibel auf ihre Umgebung reagieren können. Des Weiteren wurden Poly(L-Lactid-co-ε-Caprolacton-co-Acryloylcarbonat) und Poly(ε-Caprolacton-co-Acryloylcarbonat) (PCLAC) Copolymere hinsichtlich deren MEW Verarbeitbarkeit untersucht. Solche Kunststoffe können nach dem Druckvorgang mit UV-Strahlung radikalisch vernetzt werden und dadurch deutlich erhöhte mechanische Eigenschaften ausbilden. Für Fasern aus 90 mol.% CL und 10 mol.% AC wurden beispielsweise maximale Zugfestigkeiten von σmax = 53 ± 16 MPa ermittelt. MEW gedruckte sinusförmige Faserstrukturen aus PCLAC wiesen darüber hinaus ein biomimetisches Spannungs-Dehnung-Verhalten auf, vergleichbar zu Sehnen- und Ligamentgewebe. Eine Untersuchung der Zellviabilität von L929 murinen Fibroblasten im Eluattest, sowie eine lebend/tot-Färbung von humanen mesenchymalen Stammzellen auf den Scaffolds, ergab vielversprechende Resultate und damit ein relevantes Anwendungspotential solcher Strukturen als Implantat. Neben genannten Vorteilen, weist MEW als Verfahren bislang allerdings geringe Produktionsgeschwindigkeiten auf. Diese sind daher in den Fokus aktueller Forschungsvorhaben gerückt. Einen Ansatz hierfür bieten Spinndüsen mit hohem Innendurchmesser und erhöhter Austragsrate, wobei die optimierte elektrische Feldstärke, sowie ein Verstrecken durch die Kollektorbewegung, zu den erwünschten dünnen Fasern führen können. Dadurch kann die abwärtslimitierte Düsengröße des FDM Verfahrens überwunden werden. Visionär gedacht, könnte eine solche Anlage direkt in Krankenhäusern zur Fertigung von patienten- und defektspezifischen Implantaten eingesetzt werden. Darüber hinaus ermöglicht die hohe Präzision, zusammen mit dem Drucken von Mikro-Fasern, einen technischen Einsatz zur Herstellung von Membranen, Filtern oder funktionalen Oberflächenbeschichtungen

This thesis presents a melt electrospinning technique to fabricate highly porous and controllable poly (ε-caprolactone) (PCL) microfibers for tissue engineering applications and rehabilitation applications. Electrospinning without solvents via melt methods may be an attractive approach to tissue engineering of cell constructs where solvent accumulation or toxicity is an issue. This method is also able to produce microfibers with controllable parameters. However, the fiber diameters resulting from melt electrospinning processes are relatively large when compared to the fibers from solution electrospinning. The typical microfiber diameter from melt electrospinning was reported to be approximately 0.1mm. In order to further develop the melt electrospinning technique, we focused on the design of a melt electrospinning setup based on numerical analysis using the Solidworks 2013 simulation package and practically established a melt electrospinning setup and thermal control system for accurate experiments. One of main purposes of this thesis is the build-up of mathematical modeling to control and predict the electrospun microfiber via a more intricate understanding of their parameters such as the nozzle diameter, applied voltage, distance between the nozzle and counter electrode, temperature, flow rate, linear transitional speed, among others. The model is composed of three parts: 1) melt electrospinning process modeling, 2) fibrous helix movement modeling, and 3) build-up of microfibers modeling. The melt electrospinning process model describes an electric field, the shape of jet’s continuously changing shape, and how the polymer melt is stretched into a Taylor cone and a straight jet. The fibrous helix movement model describes movement of electrospun microfibers influenced by Lorentz force, which moves along the helix pattern. Lastly, the build-up microfiber modeling describes the accumulation of the extruded microfibers on both flat and round counter electrodes based on the physical forces involved. These models are verified by experimental data from our own customized melt electrospinning setup. Moreover, the fabricated scaffolds are tested by seeding neural progenitors derived from murine R1 embryonic stem cell lines and it demonstrates the potential of scaffolds for tissue engineering applications. To increase cell attachment and proliferation, highly porous microfibers are fabricated by combination of melt electrospinning and particulate leaching technique. Finally, auxetic stretchable PCL force sensors are fabricated by melt electrospinning for hand rehabilitation. These stretchable sensors can be used to measure applied external loads or displacement and are also attachable to various substrates. We have attempted to apply the sensors to real human hand in order to prove their functionality.
Graduate
jko@me.uvic.ca