Controlled drug delivery systems have been utilized to enhance the therapeutic effects of many current drugs by effectively delivering drugs in a time-dependent and repeatable manner. The ability to control the delivery of drugs, whether through sequential, instantaneous, sustained, delayed and/or enhanced release has the potential to provide effective dosing regimens with enhanced therapeutic effects for a plethora of diseases and injuries. For instance, such systems can enhance anti-tumoral responses or, alternatively, promoting tissue regeneration. The current need for organ and tissue replacement, repair and regeneration for patients is continually growing such that supply is not meeting the high demand primarily due to a paucity of donors as well as biocompatibility issues that lead to immune rejection of the transplant. To overcome this problem, scientists working in the field of tissue engineering and regenerative medicine have investigated the use of scaffolds as an alternative to transplantation. These scaffolds are designed to mimic the extracellular matrix (ECM) by providing structural support as well as promoting attachment, proliferation, and differentiation with the goal of yielding functional tissues or organs. Continued advancement and hybrid approaches using different material combinations and printing methodologies will further advance the progress of 3D printing technologies toward developing scaffolds, and other implantable drug delivery devices, capable of being utilized in the clinic. Such advancements will not only make inroads into improving structural integrity of implantable devices but will also provide platforms for controlled drug delivery from such devices. The primary focus of this thesis will be on controlled drug delivery as well as the integration of controlled drug delivery into 3D printed devices aimed at promoting tissue regeneration. We initially assessed the efficacy of a controlled drug delivery system for the treatment of cancer using on-demand, and sustained, release of an anticancer drug, doxorubicin (DOX), for the treatment of melanoma in a murine model. Using a melanoma model, we investigated the antitumor potential of combining ultrasound (US) with poly(lactic-co-glycolic acid) (PLGA) microspheres loaded with DOX. An in vitro release assay demonstrated an ability of US to affect the release kinetics of DOX from DOX-loaded PLGA microspheres by inducing a 12% increase in rate of release where this treatment resulted in synergistic tumor cell (B16-F10 melanoma cells) killing. Melanoma-bearing mice treated intratumorally with DOX (8 µg)-loaded microspheres and subjected to US treatment at the tumor site were shown to significantly extended survival compared to untreated mice or mice subjected to either treatment alone. The synergistic increase in survival of melanoma-challenged mice treated with the combination of US and DOX-loaded microspheres implicates a promising additional tool for combatting an otherwise currently incurable cancer. We then further investigated other novel control drug delivery systems which included a 3D printed device (tube) for the purposes of sequential drug delivery. 3D printed hollow alginate tubes were fabricated through co-axial bioprinting and then injected with PLGA to provide sequential release of distinct fluorescent dyes (model drugs), where fluorescein was initially released from alginate followed by the delayed release (up to 55 h) of rhodamine B in PLGA. With an alginate shell and a PLGA core, the fabricated tubes showed no cytotoxicity when incubated with the human embryonic kidney (HEK293) cell line or bone marrow stromal stem cells (BMSC). Microscale printing through two-photon polymerization (2PP) was then investigated for controlled drug delivery potential. Poly(ethylene glycol) dimethacrylate (PEGDMA) devices were fabricated using a Photonic Professional GT two-photon polymerization system while rhodamine B was homogenously entrapped inside the polymer matrix during photopolymerization. These devices were printed with varying porosity and morphology and using varying printing parameters such as slicing and hatching distance. Overall, tuning the hatching distance, slicing distance, and pore size of the fabricated devices provided control of rhodamine B release due to resulting changes in the motility of the small molecule and its access to structure edges. In general, increased spacing provided higher drug release while smaller spacing resulted in some occlusion, preventing media infiltration and thus resulting in reduced drug release. 2PP was further explored for its ability to tailor topographical cues in addition to controlled drug release. These physical cues, similar to those of the ECM, have been seen to promote differentiation. With 2PP, we explored microscale topographies with nanoscale precision, where different star size topographies were fabricated. It was observed that the smallest star size topographies differentiated human iPSCs towards the endoderm and mesoderm germ layer. Integrating the facility for controlled drug release into 3D printed devices provides a demand for constructs that not only need to fulfill their purpose of temporarily substituting for the missing tissue at the site of injury, but also providing the necessary cues to promote appropriate tissue regeneration. With 3D printing technology, novel drug delivery constructs were fabricated and tested to appraise functionality such as the ability to control drug delivery and the ability to function as a non-toxic medium for cellular attachment, proliferation, and forced differentiation.

Understanding cell migration is a key step in unraveling many physiological phenomena and predicting several pathologies, such as cancer metastasis. In particular, confinement has been proven to be a key factor in the cellular migration strategy choice. As our insight in the field improves, new tools are needed in order to empower biologists’ analysis capabilities. In this framework, microfluidic devices have been used to engineer the mechanical and spatial stimuli and to investigate cellular migration response in a more controlled way. In this work, we will review the existing technologies employed in the realization of microfluidic cellular migration assays, namely the soft lithography of PDMS and hydrogels and femtosecond laser micromachining. We will give an overview of the state of the art of these devices, focusing on the different geometrical configurations that have been exploited to study specific aspects of cellular migration. Our scope is to highlight the advantages and possibilities given by each approach and to envisage the future developments in in vitro migration studies under spatial confinement in microfluidic devices.

<p>  </p> <p>With the advent of femtosecond lasers in the early 1990s, ultrafast laser processing has proven to be an imperative tool for micro/nanomachining. Two-photon lithography (TPL) is one such unique microfabrication technique exploiting the nonlinear dependency of the polymerization rate on the irradiating light intensity to produce true three-dimensional structures with feature sizes beyond the diffraction limit. This characteristic has revolutionized laser material processing for the fabrication of micro and nanostructures. This research first gives a general overview of TPL, including its operating principle, experimental setup, compatible materials, and techniques for improving the final resolution of the fabricated structure. Insights to improve throughput and speed of fabrication to pave a way for the industrialization of this technique are provided.</p> <p>Following that, the report delves into the process of fabricating two true three-dimensional mechanical metamaterials via the stereolithography technique. This chapter encompasses the design, fabrication, and experimental analysis of a three-dimensional axisymmetric structure with elliptical perforations distributed periodically on the walls of the structure with varying thicknesses. Furthermore, this study discusses the significance of the elliptical perforations in terms of auxetic behavior and load-bearing capacity against a so-called plain structure under quasistatic compression loading.</p> <p>Finally, the report explores the technique of fabricating a true three-dimensional cylindrical auxetic structure via two-photon polymerization. This section of the report examines the optical setup utilized, the sample preparation procedure, and calibration experiments performed in order to fabricate a three-dimensional thin-walled right cylinder with bowtie like perforations arranged on the walls to promote the exhibition of symmetric negative Poisson’s ratio under uniaxial quasistatic compression loading.</p>

<p> 3D printing has established itself as a critical tool for manufacturing in all areas. It has evolved from a purely rapid prototyping technique into a feasible process for large-scale processing. A wide variety of 3D printing processes exist across an extreme range of size, from meters to nanometers. Much of the current technological advances come from pushing fabrication techniques to smaller and smaller scales. For 3D printing this has led to the rise of two-photon polymerization, a direct laser writing process with submicron structuring capabilities. Two-photon polymerization has proven its worth as a nanoscale 3D fabrication technique but is often considered slow and expensive, two undesirable qualities for high throughput manufacturing. Parallelization methods such as projection lithography are potential solutions to increasing the throughput capabilities of two-photon polymerization 3D printing. Additionally, the drive for further reducing the print size has inspired printing resolution enhancing strategies in two-photon polymerization printing by processes such as stimulated emission depletion (STED) and other STED-inspired pathways. This work will explore avenues for improving two-photon polymerization printing throughput and resolution.</p> <p> First, a two-photon polymerization printing system is constructed with a secondary laser for controlling polymerization inhibition. Through a STED process, a 65 nm wide printed line feature was achieved. Alongside this, a characterization and verification methodology for choosing new photoinitiator molecules for similar inhibition lithography processes is presented. Through implementation of tests such as Z-scan, fluorescence depletion, ultrafast transient spectroscopy and UV-Vis absorption and fluorescence measurements a promising new photoinitiator with 5-factor improvement in printing efficiency is found. </p> <p> Second, a projection lithography scheme is developed for rapid two-photon 3D printing. A digital micro-mirror device (DMD) is utilized for dynamic pattern generation and the effects of its dispersion properties are considered. Through a spatiotemporal focusing process, continuous 3D printing is achieved at vertical prints speeds of 1 mm s-1. Simulations performed representing this rapid printing process indicate a ~1 µm layer print feature size for large areas of exposure. Comparably, a printed vertical feature size of ~ 1 µm was achieved. Lateral feature sizes ~200 nm were also demonstrated in fabrication. A variety of complex 3D structures are printed for demonstration of the spatiotemporal focusing projection lithography process including millimeter scale objects with micrometer scale 3D features.</p> <p> Finally, resolution enhancing strategies are implemented into the continuous, projection two-photon lithography technique. An investigation of the inhibition properties of a variety of photoinitiator systems for inhibiting polymerization achieved with low repetition rate laser exposure is presented. A planar polymerization inhibiting region is generated by creating a light sheet propagating perpendicularly to the projection printing plane. </p>

Os micro-ressonadores que suportam whispering gallery modes têm atraído a atenção da comunidade científica devido a sua grande capacidade de confinar a luz, propriedade que faz dessas estruturas plataformas ideais para o desenvolvimento de pesquisa fundamental como interação da radiação com a matéria e óptica não linear. Além disso, suas características como operação em frequências do visível e de telecomunicações, facilidade de integração e alta sensitividade os tornam extremamente flexíveis para aplicações que vão desde filtros ópticos até sensores. Neste trabalho, demonstramos a fabricação de tais micro-ressonadores via fotopolimerização por absorção de dois fótons (FA2F). Esta técnica apresenta uma série de vantagens para a confecção de micro-dispositivos, sendo elas a capacidade de resolução inferior ao limite de difração, a flexibilidade de formas e ainda, a possibilidade de incorporar compostos de interesse à matriz polimérica a fim de introduzir novas funcionalidades ao material que compõe a estrutura final. Ademais, diferentes polímeros podem ser utilizados para a fabricação das microestruturas, tornando a técnica viável para uma vasta gama de aplicações. As microestruturas poliméricas que fabricamos são micro-cilindros ocos de boa integridade estrutural com 45 &mu;m de diâmetro externo e 100 nm de rugosidade de superfície, o que as torna potencialmente aplicáveis como micro-ressonadores para frequências de operação típicas de telecomunicações. A fim de acoplar luz nessas estruturas, em colaboração com a Universidade de Valência, na Espanha, montamos um aparato de acoplamento. Neste aparato, a luz proveniente de uma fonte de luz centrada em 1540 nm é acoplada nos micro-ressonadores via campo evanescente por meio do uso de uma fibra óptica estirada de 1.5 &mu;m de diâmetro. A potência transmitida é guiada para um analisador de espectro óptico, onde é possível identificar os modos ressonantes, representados como picos de atenuação com free spectral range em torno de 9.8 nm. Ao término desse projeto, um aparato similar foi montado no Grupo de Fotônica do IFSC/USP, a partir do qual pudemos medir os modos ressonantes tanto de fibras ópticas estiradas quanto dos micro-cilindros poliméricos. A finesse dos micro-ressonadores poliméricos caracterizados varia de 2.51 a 4.35, sendo da mesma ordem de grandeza do valor reportado na literatura para ressonadores de alta performance fabricados por FA2F a partir da mesma formulação de resina polimérica que utilizamos. / Whispering gallery modes microresonators have been attracting increasing interest due to their ability to strongly confine light within small dielectric volumes. This property is quite useful for basic research involving light-matter interaction and nonlinear optics, but their applications go beyond that. The ease of fabrication, on-chip integration and operation at telecommunication frequencies make them suitable for a variety of practical applications, including photonic filters and sensing. In the current work, we demonstrate the fabrication of such resonators via two-photon polymerization. Using this technique, complex 3D structures with submicrometer feature size can be produced. Besides, the flexibility of geometry and the possibility of incorporating a variety of additional materials, such as organic compounds make it a powerful tool for the fabrication of microresonators. The microstructures we have fabricated are 45 &mu;m outer diameter hollow microcylinders, with good structural integrity and sidewall roughness estimated in 100 nm, which make their application as microresonators feasible in the near infrared wavelength regime. In order to couple light within these microresonators, an experimental setup was built at University of Valencia to implement the coupling. In this setup, light from a 1540 nm-centered broadband source was coupled into the fabricated microresonators via evanescent field using a 1.5 &mu;m waist tapered fiber. The transmitted light was then guided to an optical spectral analyzer, where it was possible to measure resonances, represented as attenuation peaks, with free spectral range of about 9.8 nm. Afterwards, a similar experimental setup was assembled in the Photonics group at IFSC/USP, where we could observe resonances of both tapered optical fibers and the polymeric microresonators fabricated by means of two-photon polymerization. The finesse of the polymeric microresonators was estimated in 4.35, being in the same order of the finesse reported in the literature for high performance microring resonators fabricated using the same polymeric resin.

Os micro-ressonadores que suportam whispering gallery modes têm atraído a atenção da comunidade científica devido a sua grande capacidade de confinar a luz, propriedade que faz dessas estruturas plataformas ideais para o desenvolvimento de pesquisa fundamental como interação da radiação com a matéria e óptica não linear. Além disso, suas características como operação em frequências do visível e de telecomunicações, facilidade de integração e alta sensitividade os tornam extremamente flexíveis para aplicações que vão desde filtros ópticos até sensores. Neste trabalho, demonstramos a fabricação de tais micro-ressonadores via fotopolimerização por absorção de dois fótons (FA2F). Esta técnica apresenta uma série de vantagens para a confecção de micro-dispositivos, sendo elas a capacidade de resolução inferior ao limite de difração, a flexibilidade de formas e ainda, a possibilidade de incorporar compostos de interesse à matriz polimérica a fim de introduzir novas funcionalidades ao material que compõe a estrutura final. Ademais, diferentes polímeros podem ser utilizados para a fabricação das microestruturas, tornando a técnica viável para uma vasta gama de aplicações. As microestruturas poliméricas que fabricamos são micro-cilindros ocos de boa integridade estrutural com 45 &mu;m de diâmetro externo e 100 nm de rugosidade de superfície, o que as torna potencialmente aplicáveis como micro-ressonadores para frequências de operação típicas de telecomunicações. A fim de acoplar luz nessas estruturas, em colaboração com a Universidade de Valência, na Espanha, montamos um aparato de acoplamento. Neste aparato, a luz proveniente de uma fonte de luz centrada em 1540 nm é acoplada nos micro-ressonadores via campo evanescente por meio do uso de uma fibra óptica estirada de 1.5 &mu;m de diâmetro. A potência transmitida é guiada para um analisador de espectro óptico, onde é possível identificar os modos ressonantes, representados como picos de atenuação com free spectral range em torno de 9.8 nm. Ao término desse projeto, um aparato similar foi montado no Grupo de Fotônica do IFSC/USP, a partir do qual pudemos medir os modos ressonantes tanto de fibras ópticas estiradas quanto dos micro-cilindros poliméricos. A finesse dos micro-ressonadores poliméricos caracterizados varia de 2.51 a 4.35, sendo da mesma ordem de grandeza do valor reportado na literatura para ressonadores de alta performance fabricados por FA2F a partir da mesma formulação de resina polimérica que utilizamos. / Whispering gallery modes microresonators have been attracting increasing interest due to their ability to strongly confine light within small dielectric volumes. This property is quite useful for basic research involving light-matter interaction and nonlinear optics, but their applications go beyond that. The ease of fabrication, on-chip integration and operation at telecommunication frequencies make them suitable for a variety of practical applications, including photonic filters and sensing. In the current work, we demonstrate the fabrication of such resonators via two-photon polymerization. Using this technique, complex 3D structures with submicrometer feature size can be produced. Besides, the flexibility of geometry and the possibility of incorporating a variety of additional materials, such as organic compounds make it a powerful tool for the fabrication of microresonators. The microstructures we have fabricated are 45 &mu;m outer diameter hollow microcylinders, with good structural integrity and sidewall roughness estimated in 100 nm, which make their application as microresonators feasible in the near infrared wavelength regime. In order to couple light within these microresonators, an experimental setup was built at University of Valencia to implement the coupling. In this setup, light from a 1540 nm-centered broadband source was coupled into the fabricated microresonators via evanescent field using a 1.5 &mu;m waist tapered fiber. The transmitted light was then guided to an optical spectral analyzer, where it was possible to measure resonances, represented as attenuation peaks, with free spectral range of about 9.8 nm. Afterwards, a similar experimental setup was assembled in the Photonics group at IFSC/USP, where we could observe resonances of both tapered optical fibers and the polymeric microresonators fabricated by means of two-photon polymerization. The finesse of the polymeric microresonators was estimated in 4.35, being in the same order of the finesse reported in the literature for high performance microring resonators fabricated using the same polymeric resin.

Printing of 3-dimensional nanostructures with high-resolution by two-photon polymerization has gained significant attention recently. Isopropyl thioxanthone (ITX) has been studied and used as a photoinitiator because of its unique property in initiating and depleting polymerization, but to further improve the resolution of 3D structures, new photoinitiating materials are necessary to decrease the power requirements especially in industrial world. In this dissertation, different new types of thioxanthone-based photoinitiators were synthesized and our new initiators possessed a clear enhancement in terms of excitation over ITX. To clearly reveal the writing mechanism behind it, the behavior of the initiators was evaluated by several methods such as low temperature phosphorescence spectroscopy and density functional theory (DFT) calculations. The first type of new molecules with alkyne bridge will be discussed in chapter 2 and the further developed initiators with electron donating and withdrawing groups will be discussed in chapter 3. By modifying the structure of ITX, we have revealed and proposed an important pathway to guide future development of photoinitiators in direct laser writing.

The study of cellular migration dynamics and strategies plays a relevant role in the understanding of both physiological and pathological processes. An important example could be the link between cancer cell motility and tumor evolution into metastatic stage. These strategies can be strongly influenced by the extracellular environment and the consequent mechanical constrains. In this framework, the possibility to study the behavior of single cells when subject to specific topological constraints could be an important tool in the hands of biologists. Two-photon polymerization is a sub-micrometric additive manufacturing technique that allows the fabrication of 3D structures in biocompatible resins, enabling the realization of ad hoc biochips for cell motility analyses, providing different types of mechanical stimuli. In our work, we present a new strategy for the realization of multilayer microfluidic lab-on-a-chip constructs for the study of cell motility which guarantees complete optical accessibility and the possibility to freely shape the migration area, to tailor it to the requirements of the specific cell type or experiment. The device includes a series of micro-constrictions that induce different types of mechanical stress on the cells during their migration. We show the realization of different possible geometries, in order to prove the versatility of the technique. As a proof of concept, we present the use of one of these devices for the study of the motility of murine neuronal cancer cells under high physical confinement, highlighting their peculiar migration mechanisms.

Η διφωτονική απορρόφηση (ΔΦΑ) είναι το μη γραμμικό-φαινόμενο κατά το οποίο δύο φωτόνια απορροφούνται ταυτόχρονα σε ένα υλικό μέσο. Τα τελευταία 20 χρόνια το φαινόμενο αυτό έχει προσελκύσει ιδιαίτερα το ενδιαφέρον της επιστημονικής κοινότητας για την ανακάλυψη νέων αποδοτικών ενώσεων λόγω και των εφαρμογών που έχει βρεθεί ότι μπορεί να προσφέρει. Η διφωτονική μικροσκοπία, ο διφωτονικός πολυμερισμός, η τρισδιάστατη αποθήκευση δεδομένων είναι μερικές από τις σημαντικές εφαρμογές που εκμεταλλεύονται τα πλεονεκτήματα της ΔΦΑ. Τη τελευταία δεκαετία η έρευνα επικεντρώνεται στη θεωρητική και πειραματική μελέτη της ΔΦΑ οργανικών ενώσεων με σκοπό την διασύνδεσης των χαρακτηριστικών της δομής των μορίων με την ΔΦΑ που παρέχουν και την εύρεση στρατηγικών σύνθεσης οργανικών ενώσεων υψηλής ΔΦΑ. Επίσης πολλές είναι οι επιστημονικές εργασίες που χρησιμοποιούν τις ενώσεις αυτές στην έρευνα για την ανάπτυξη και βελτίωση των διφωτονικών εφαρμογών. Στη διατριβή αυτή γίνεται μελέτη της ΔΦΑ συζυγιακών οργανικών ενώσεων αποτελούμενες από ένα κεντρικό τμήμα συνδεδεμένο αντιδιαμετρικά με δυο ίδιους υποκαταστάτες. Τα συμμετρικά αυτά μόρια έχουν ως κεντρικό τμήμα φλουορένιο ή αλκόξυ-φαινυλένιο ή καρβοξύλιο και διάφορους υποκαταστάτες στα άκρα. Η τεχνική μέτρησης διφωτονικά διεγερμένου φθορισμού με laser femtosecond χρησιμοποιήθηκε για τον υπολογισμό των τιμών της ενεργούς διατομής ΔΦΑ των μορίων στη φασματική περιοχή 750-840nm. Από τη συγκριτική μελέτη των ενώσεων προκύπτει ότι ο υποκαταστάτης φθαλιμίδιο προσφέρει την μεγαλύτερη ΔΦΑ και για τα τρία διαφορετικά κεντρικά τμήματα, ενώ όταν συνδέεται με το φλουορένιο έχει την υψηλότερη ενεργό διατομή ΔΦΑ φθάνοντας την τιμή των 1650GM στη φασματική περιοχή που μελετήθηκε. Επίσης παρατηρήθηκε ότι ο υποκαταστάτης ναφθαλιμίδιο προκαλεί μετατόπιση του μεγίστου ΔΦΑ σε αρκετά μεγαλύτερα μήκη κύματος. Παρόμοια μετατόπιση παρατηρείται και για τα μόρια με κεντρικό τμήμα το καρβαζόλιο. Η μετατόπιση αυτή του φάσματος ΔΦΑ δεν αντιστοιχεί σε μετατόπιση του φάσματος μονοφωτονικής απορρόφησης. Οφείλεται στην άρση της κεντροσυμμετρίας του μορίου, λόγω της διαμόρφωσης της δομής του, η οποία κάνει επιτρεπτή τη διφωτονική μετάβαση στην πρώτη μονοφωτονικά επιτρεπτή ενεργειακή κατάσταση που ειδάλλως είναι απαγορευμένη. Η επίδραση του διαλύτη στις διφωτονικές ιδιότητες των μορίων μελετάται επίσης στη διατριβή αυτή. Για τις τέσσερις πιο αποδοτικές ενώσεις, ως προς το διφωτονικά διεγερμένο φθορισμό, έγιναν μετρήσεις για τον υπολογισμό των φασμάτων ΔΦΑ τους σε πέντε διαλύτες διαφορετικής πολικότητας (διηλεκτρικής σταθεράς). Από τις μετρήσεις φάνηκε η σημαντική επίδραση του διαλύτη στη ΔΦΑ των τεσσάρων χρωστικών η οποία μεγιστοποιείται σε διαλύτη μεσαίας πολικότητας, ο τρεις στην ακετοφαινόνη και μια στο THF. Η χαμηλή κβαντική απόδοση φθορισμού που παρατηρείται στα διαλύματα με ακετοφαινόνης μειώνει αρκετά το διφωτονικά προκαλούμενο φθορισμό και κατατάσσει τo THF ως τον αποδοτικότερο διαλύτη για διφωτονικές εφαρμογές που εκμεταλλεύονται το φθορισμό. Επίσης και ο μη πολικός διαλύτης τολουόλιο προκαλεί έντονο φθορισμό λόγω της υψηλής κβαντικής απόδοσης παρόλο που επιφέρει δραστική μείωση της ΔΦΑ σε σύγκριση με τους άλλους διαλύτες που μελετήθηκαν. Δύο από τις χρωστικές που μελετήθηκαν ως προς τη ΔΦΑ τους χρησιμοποιήθηκαν ως φωτοεκκινητής πολυμερισμού ενός ακρυλικού μονομερούς και την μελέτη του πολυμερισμού συναρτήσει διαφόρων παραμέτρων ακτινοβόλησης της ρητίνης. Οι χρωστικές αυτές έχουν φλουορένιο ως κεντρικό τμήμα και υποκαταστάτες στα άκρα φθαλιμίδιο ή τριφαινυλαμίνη. Και οι δύο χρωστικές δύναται να προκαλέσουν τον πολυμερισμό του μονομερούς με ακτινοβόληση υπερβραχέων παλμών στα 800nm, αλλά η προσθήκη αμίνης ως συνεκκινητή μειώνει το κατώφλι ισχύος εκκίνησης του. Επίσης, σε υψηλές τιμές ισχύος ακτινοβόλησης στο ίδιο μήκος κύματος παρατηρήθηκε αυτο-πολυμερισμός του μονομερούς γεγονός το οποίο έχει επισημανθεί μία μόνο ακόμη φορά σε μια εργασία με ρητίνη μίξης τριών ακρυλικών μονομερών. Δύο ρητίνες που παρασκευάστηκαν στην παρούσα διατριβή αναμιγνύοντας το μονομερές με τη κάθε χρωστική και με προσθήκη του συνεκκινητή μελετήθηκαν για την εξάρτηση του πολυμερισμού από τον φακό εστίασης και από την ισχύ, την ταχύτητα σάρωσης και τη διάμετρο της δέσμης. Για το σκοπό αυτό κατασκευάστηκαν δοκίμια στα οποία εγγράφονταν πολυμερισμένες γραμμές μεταβάλλοντας κάποια από τις παραπάνω παραμέτρους και μετρώντας το πάχος και το ύψος των γραμμών αυτών από τις εικόνες μικροσκοπίου σάρωσης ηλεκτρονίων. Το γεγονός ότι ο μικρότερης ισχύος φακός παρατηρηθηκε ότι προκαλεί λεπτότερη γραμμή εξηγείται από το φαινόμενο κατωφλίου του πολυμερισμού. Ωστόσο η λεπτότερη γραμμή επιτυγχάνεται με τον ισχυρότερο φακό καθώς μπορεί να μειωθεί περισσότερο η ισχύς της δέσμης λόγω του χαμηλότερου κατωφλίου εκκίνησης του πολυμερισμού για το φακό αυτό. Η αύξηση της ταχύτητας σάρωσης ή η μείωση της ισχύος της δέσμης επιφέρουν μικρότερες διαστάσεις πάχους και ύψους της γραμμής που πολυμερίζεται. Επίσης η κατανομή της ισχύος ακτινοβόλησης σε ολόκληρο το πίσω άνοιγμα του φακού επιφέρει καλύτερη εστίαση και λεπτότερη πολυμερισμένη γραμμή. Ωστόσο η κατάλληλη χωρική διαμόρφωση της δέσμης μπορεί να μειώσει τις διαστάσεις των πολυμερισμένων δομών. / -

In the last decades, additive manufacturing (AM), also called three-dimensional (3D) printing, has advanced micro/nano-fabrication technologies, especially in applications like lightweight engineering, optics, energy, and biomedicine. Among these 3D printing technologies, two-photon polymerization (TPP) offers the highest resolution (even at the nanometric scale), reproducibility and the possibility to create monolithically 3D complex structures with a variety of materials (e.g. organic and inorganic, passive and active). Such active materials change their shape upon an applied stimulus or degrade over time at certain conditions making them dynamic and reconfigurable (also called 4D printing). This is particularly interesting in the field of medical microrobotics as complex functions such as gentle interactions with biological samples, adaptability when moving in small capillaries, controlled cargo-release profiles, and protection of the encapsulated cargoes, are required. Here we review the physics, chemistry and engineering principles of TPP, with some innovations that include the use of micromolding and microfluidics, and explain how this fabrication schemes provide the microrobots with additional features and application opportunities. The possibility to create microrobots using smart materials, nano- and biomaterials, for in situ chemical reactions, biofunctionalization, or imaging is also put into perspective. We categorize the microrobots based on their motility mechanisms, function, and architecture, and finally discuss the future directions of this field of research.