Development of a polyplexes-based miRNA delivery system in a 3D-bioplotted osteoarthritis cellular model.

L'osteoartrite (OA) è una patologia infiammatorio/degenerativa ossea per la quale non sono disponibili terapie causali efficaci ma solo approcci palliativi per la riduzione del dolore cronico. E’ quindi giustificato un investimento per individuare nuove strategie di trattamento. In quest’ottica, lo scopo di questa tesi è stato quello di indagare l’efficacia di polyplexi a base di chitosano o di PEI-g-PEG in un modello cellulare 3D in vitro basato su un hydrogel di Gellan Gum Metacrilato (GGMA) con a bordo condrociti in condizioni simulate di OA. Inizialmente sono state studiate la dimensione e il potenziale-Z di un pool di formulazioni di poliplexi. Quindi se ne è valutata la citocompatibilità utilizzando cellule staminali mesenchimali immortalizzate Y201. Infine, una miscela di GGMA, cellule e polyplexi è stata utilizzata per la stampa 3D di campioni che sono stati coltivati fino a 14 giorni. La condizione OA è stata simulata trattando le cellule con una miscela di citochine implicate nello sviluppo della malattia. Tutte le formulazioni a base di chitosano e due basate su PEI-g-PEG si sono dimostrate citocompatibili e sono hanno veicolato i miRNA nelle cellule (come mostrato dai risultati di analisi in fluorescenza). I risultati delle colorazioni H&E e AlcianBlue hanno confermato che il terreno condizionato ha ben ricreato le condizioni di OA. I polyplexi a base di chitosano e PEI-g-PEG hanno controbilanciato gli effetti delle citochine. Risultati incoraggianti, anche se da approfondire ulteriormente, provengono anche dall’analisi di espressione (RT-PCR) di cinque geni specifici della cartilagine. Concludendo, questo modello ha ben riprodotto le condizioni di OA in vitro; il chitosano ha mostrato di essere un adeguato veicolo per un trattamento a base di miRNA; il PEI-g-PEG si propone come un'alternativa più economica e ragionevolmente affidabile, sebbene il rischio di citotossicità alle concentrazioni più elevate richieda una più esteva validazione sperimentale.

Characterization of a 3D-printed low-cost flexible dielectric material for the realization of innovative WPT wearable applications

The aim of this thesis is to demonstrate that 3D-printing technologies can be considered significantly attractive in the production of microwave devices and in the antenna design, with the intention of making them lightweight, cheaper, and easily integrable for the production of wireless, battery-free, and wearable devices for vital signals monitoring. In this work, a new 3D-printable, low-cost resin material, the Flexible80A, is proposed as RF substrate in the implementation of a rectifying antenna (rectenna) operating at 2.45 GHz for wireless power transfer. A careful and accurate electromagnetic characterization of the abovementioned material, revealing it to be a very lossy substrate, has paved the way for the investigation of innovative transmission line and antenna layouts, as well as etching techniques, possible thanks to the design freedom enabled by 3D-printing technologies with the aim of improving the wave propagation performance within lossy materials. This analysis is crucial in the design process of a patch antenna, meant to be successively connected to the rectifier. In fact, many different patch antenna layouts are explored varying the antenna dimensions, the substrate etchings shape and position, the feeding line technology, and the operating frequency. Before dealing with the rectification stage of the rectenna design, the hot and long-discussed topic of the equivalent receiving antenna circuit representation is addressed, providing an overview of the interpretation of different authors about the issue, and the position that has been adopted in this thesis. Furthermore, two rectenna designs are proposed and simulated with the aim of minimizing the dielectric losses. Finally, a prototype of a rectenna with the antenna conjugate matched to the rectifier, operating at 2.45 GHz, has been fabricated with adhesive copper on a substrate sample of Flexible80A and measured, in order to validate the simulated results.

Conceptualization of an Electric Compact SUV for the American and Italian Market using the IDeS Method

Electric cars are increasingly popular due to a transition of mobility towards more sustainable forms. From an increasingly green and pollution reduction perspective, there are more and more incentives that encourage customers to invest in electric cars. Using the Industrial Design and Structure (IDeS) research method, this project has the aim to design a new electric compact SUV suitable for all people who live in the city, and for people who move outside urban areas. In order to achieve the goal of developing a new car in the industrial automotive environment, the compact SUV segment was chosen because it is a vehicle very requested by the costumers and it is successful in the market due to its versatility. IDeS is a combination of innovative and advanced systematic approaches used to set up a new industrial project. The IDeS methodology is sequentially composed of Quality Function Deployment (QFD), Benchmarking (BM), Top-Flop analysis (TFA), Stylistic Design Engineering (SDE), Design for X, Prototyping, Testing, Budgeting, and Planning. The work is based on a series of steps and the sequence of these must be meticulously scheduled, imposing deadlines along the work. Starting from an analysis of the market and competitors, the study of the best and worst existing parameters in the competitor’s market is done, arriving at the idea of a better product in terms of numbers and innovation. After identifying the characteristics that the new car should have, the other step is the styling part, with the definition of the style and the design of the machine on a 3D CAD. Finally, it switches to the prototyping and testing phase to see if the product is able to work. Ultimately, intending to place the car on the market, it is essential to estimate the necessary budget for a possible investment in this project.

Caratterizzazione meccanica ad alta temperatura dell'acciaio inossidabile AISI 316L prodotto con processo additivo

Analisi meccanica e microstrutturale a temperatura ambiente e alta temperatura (400 °C) dell'acciaio inossidabile AISI 316L prodotto tramite additive manufacturing (Laser Powder Bed Fusion) e rivestito con un sistema multistrato composto da nichelatura al fosforo e diamond like carbon. Il materiale è stato testato meccanicamente tramite prove di trazione, fatica a basso numero di cicli e durezza. Queste hanno dimostrato come il rivestimento non modifica significativamente le proprietà a trazione del materiale (sia a 20°C che a 400 °C) mentre allunga notevolmente la vita a fatica del provino a temperatura ambiente. Le analisi di durezza hanno evidenziato il diverso comportamento del rivestimento a temperatura ambiente e a 400 °C in quanto, alla temperatura di 300 °C, subisce una ricristallizzazione che ne determina un indurimento. L'analisi microstrutturale ha previsto l'osservazione del materiale al microscopio ottico ed elettronico a scansione FEG-SEM, la quale ha permesso di identificare i melt pool, grani colonnari e struttura cellulare che compongono il materiale. La struttura cellulare scompare a 400 °C mentre i melt pool ed i grani colonnari restano invariati. Oltre a queste analisi sono state svolte anche delle osservazioni per quanto riguarda le superfici di frattura del materiale le quali hanno mostrato una tipologia di frattura duttile nei provini di trazione mentre i campioni a fatica presentano una tipica frattura a fatica, ovvero che comprende un innesco, una zona di propagazione della cricca ed una zona di rottura di schianto. I campioni rivestiti presentavano l'innesco nella zona interfacciale tra substrato e rivestimento a causa delle tensioni residue, nonostante l'adesione del rivestimento con l'acciaio fosse ottimale.

Studio di fabbricabilità per strutture porose interconnesse con macchina PolyJet

Il presente caso di studio prende forma da una problematica su una metodologia di stampa 3D relativamente nuova, la tecnologia PolyJet. Nella tesi, si verificano ed osservano i limiti di fabbricazione dei modelli ottenuti da questo processo di stampa, dovuti al particolare materiale di supporto che utilizza la tecnologia medesima a differenza di altre tecnologie esistenti. Nei dettagli, si è deciso di stampare dei particolari provini costituiti da strutture porose interconnesse, ottenute tramite superfici TPMS (Triply Periodical Minimal Surfaces), caratterizzate da fori molto piccoli e profondi ed osservare la facilità o la difficoltà nel rimuovere il materiale di supporto da ognuno di essi attraverso le attrezzature disponibili in laboratorio. I risultati ottenuti dall’esperimento hanno mostrato che, riducendo sempre di più i fori delle strutture dei provini, si va a complicare l’azione di rimozione del materiale di supporto, rendendo il processo difficile da compiere come nell’ultimo provino realizzato. Si evince che, nei tre casi osservati, è possibile rimuovere tutto il materiale di supporto dalle strutture, anche nell’ultimo manufatto, con fori di dimensione di un millimetro.

A new concept for a Sport-Coupè car developed with the Industrial Design Structure method (IDeS)

The aim of this study, conducted in collaboration with Lawrence Technological University in Detroit, is to create, through the method of the Industrial Design Structure (IDeS), a new concept for a sport-coupe car, based on a restyling of a retro model (Ford Mustang 1967). To date, vintage models of cars always arouse great interest both for the history behind them and for the classic and elegant style. Designing a model of a vehicle that can combine the charm of retro style with the innovation and comfort of modern cars would allow to meet the needs and desires of a large segment of the market that today is forced to choose between past and future. Thanks to a well-conceived concept car an automaker company is able to express its future policy, to make a statement of intent as, such a prototype, ticks all the boxes, from glamour and visual wow-factor to technical intrigue and design fascination. IDeS is an approach that makes use of many engineering tools to realize a study developed on several steps that must be meticulously organized and timed. With a deep analysis of the trends dominating the automotive industry it is possible to identify a series of product requirements using quality function deployment (QFD). The considerations from this first evaluation led to the definition of the technical specifications via benchmarking (BM) and top-flop analysis (TFA). Then, the structured methodology of stylistic design engineering (SDE) is applied through six phases: (1) stylistic trends analysis; (2) sketches; (3) 2D CAD drawings; (4) 3D CAD models; (5) virtual prototyping; (6) solid stylistic model. Finally, Developing the IDeS method up to the final stages of Prototypes and Testing you get a product as close as possible to the ideal vehicle conceptualized in the initial analysis.

Freeform fabrication of aluminum metal-matrix composites

A series of metal-matrix composites were formed by extrusion freeform, fabrication of a sinterable aluminum alloy in combination with silicon carbide particles and whiskers, carbon fibers, alumina particles, and hollow flyash cenospheres. Silicon carbide particles were most successful in that the composites retained high density with up to 20 vol% of reinforcement and the strength approximately doubles over the strength of the metal matrix alone. Comparison with simple models suggests that this unexpectedly high degree of reinforcement can be attributed to the concentration of small silicon carbide particles around the larger metal powder. This fabrication method also allows composites to be formed with hollow spheres that cannot be formed by other powder or melt methods.

Drilling damage in composite material

The characteristics of carbon fibre reinforced laminates have widened their use from aerospace to domestic appliances, and new possibilities for their usage emerge almost daily. In many of the possible applications, the laminates need to be drilled for assembly purposes. It is known that a drilling process that reduces the drill thrust force can decrease the risk of delamination. In this work, damage assessment methods based on data extracted from radiographic images are compared and correlated with mechanical test results—bearing test and delamination onset test—and analytical models. The results demonstrate the importance of an adequate selection of drilling tools and machining parameters to extend the life cycle of these laminates as a consequence of enhanced reliability.

Reforço de estruturas de betão armado com CFRP

Ao longo dos anos as estruturas existentes têm sido adaptadas para novas utilizações. No entanto, devido aos condicionalismos arquitetónicos e patrimoniais, a demolição e substituição por estruturas novas, pode-se tornar pouco viável, sendo cada vez mais exequível a opção de reforçar. A presente dissertação refere-se a uma dessas opções de reforço nomeadamente ao reforço de estruturas em betão armado com CFRP (Compósitos Reforçados com Fibras de Carbono), nomeadamente lajes e vigas. Os objetivos principais deste trabalho consistem em desenvolver uma proposta de critérios de dimensionamento de estruturas de betão armado reforçadas com CFRP tendo por base o disposto no Eurocódigo 2 comparando -a com o relatório técnico publicado “bulletin 14 - Externally bonded FRP reinforcement for RC structures”, da Fédération Internationale du Béton. Recorrendo à revisão bibliográfica, onde estão referidos temas como as características dos materiais de um sistema FRP, as suas técnicas de reforço e com uma exposição do comportamento das vigas reforçadas à flexão, particularmente no seu comportamento mecânico e modos de ruína associados a este tipo de reforço. Apresentam-se duas metodologias de cálculo para dimensionamento deste tipo de reforço para os diferentes estados limites, e aplicam-se a cada uma das metodologias de cálculo a uma viga com necessidade de reforço à flexão e ao corte, devido a um aumento de esforços provocado pelo aumento da sobrecarga. Desenvolve-se um estudo experimental onde se pretende avaliar a eficácia de um sistema de reforço à flexão com compósitos de CFRP colado externamente a uma viga e com diferentes taxas de reforço.

Self-monitoring of freeze–thaw damage using triphasic electric conductive concrete

The effect of freeze–thaw cycles on concrete is of great importance for durability evaluation of concrete structures in cold regions. In this paper, damage accumulation was studied by following the fractional change of impedance (FCI) with number of freeze–thaw cycles (N). The nano-carbon black (NCB), carbon fiber (CF) and steel fiber (SF) were added to plain concrete to produce the triphasic electrical conductive (TEC) and ductile concrete. The effects of NCB, CF and SF on the compressive strength, flexural properties, electrical impedance were investigated. The concrete beams with different dosages of conductive materials were studied for FCI, N and mass loss (ML), the relationship between FCI and N of conductive concrete can be well defined by a first order exponential decay curve. It is noted that this nondestructive and sensitive real-time testing method is meaningful for evaluating of freeze–thaw damage in concrete.

Thermal conditions affecting heat transfer in FDM/FFE: a contribution towards the numerical modelling of the process

The performance of parts produced by Free Form Extrusion (FFE), an increasingly popular additive manufacturing technique, depends mainly on their dimensional accuracy, surface quality and mechanical performance. These attributes are strongly influenced by the evolution of the filament temperature and deformation during deposition and solidification. Consequently, the availability of adequate process modelling software would offer a powerful tool to support efficient process set-up and optimisation. This work examines the contribution to the overall heat transfer of various thermal phenomena developing during the manufacturing sequence, including convection and radiation with the environment, conduction with support and between adjacent filaments, radiation between adjacent filaments and convection with entrapped air. The magnitude of the mechanical deformation is also studied. Once this exercise is completed, it is possible to select the material properties, process variables and thermal phenomena that should be taken in for effective numerical modelling of FFE.

Fused deposition modeling with polypropylene

This paper addresses the potential of polypropylene (PP) as a candidate for fused deposition modeling (FDM)-based 3D printing technique. The entire filament production chain is evaluated, starting with the PP pellets, filament production by extrusion and test samples printing. This strategy enables a true comparison between parts printed with parts manufactured by compression molding, using the same grade of raw material. Printed samples were mechanically characterized and the influence of filament orientation, layer thickness, infill degree and material was assessed. Regarding the latter, two grades of PP were evaluated: a glass-fiber reinforced and a neat, non-reinforced, one. The results showed the potential of the FDM to compete with conventional techniques, especially for the production of small series of parts/components; also, it was showed that this technique allows the production of parts with adequate mechanical performance and, therefore, does not need to be restricted to the production of mockups and prototypes.

Desenvolvimento e actualização do controlo de um equipamento de prototipagem rápida

Dissertação de mestrado integrado em Engenharia Mecânica

Current approaches and future perspectives on strategies for the development of personalized tissue engineering therapies

Personalized tissue engineering and regenerative medicine (TERM) therapies propose patient-oriented effective solutions, considering individual needs. Cell-based therapies, for example, may benefit from cell sources that enable easier autologous set-ups or from recent developments on IPS cells technologies towards effective personalized therapeutics. Furthermore, the customization of scaffold materials to perfectly fit a patientâ s tissue defect through rapid prototyping technologies, also known as 3D printing, is now a reality. Nevertheless, the timing to expand cells or to obtain functional in vitrotissue substitutes prior to implantation prevents advancements towards routine use upon patient´s needs. Thus, personalized therapies also anticipate the importance of creating off-the-shelf solutions to enable immediately available tissue engineered products. This paper reviews the main recent developments and future challenges to enable personalized TERM approaches and to bring these technologies closer to clinical applications.

High precision anatomy for MEG.

Precise MEG estimates of neuronal current flow are undermined by uncertain knowledge of the head location with respect to the MEG sensors. This is either due to head movements within the scanning session or systematic errors in co-registration to anatomy. Here we show how such errors can be minimized using subject-specific head-casts produced using 3D printing technology. The casts fit the scalp of the subject internally and the inside of the MEG dewar externally, reducing within session and between session head movements. Systematic errors in matching to MRI coordinate system are also reduced through the use of MRI-visible fiducial markers placed on the same cast. Bootstrap estimates of absolute co-registration error were of the order of 1mm. Estimates of relative co-registration error were <1.5mm between sessions. We corroborated these scalp based estimates by looking at the MEG data recorded over a 6month period. We found that the between session sensor variability of the subject's evoked response was of the order of the within session noise, showing no appreciable noise due to between-session movement. Simulations suggest that the between-session sensor level amplitude SNR improved by a factor of 5 over conventional strategies. We show that at this level of coregistration accuracy there is strong evidence for anatomical models based on the individual rather than canonical anatomy; but that this advantage disappears for errors of greater than 5mm. This work paves the way for source reconstruction methods which can exploit very high SNR signals and accurate anatomical models; and also significantly increases the sensitivity of longitudinal studies with MEG.