Recentemente, utilizando a técnica de auto-agregação (\"self-assembly\") foi possível visualizar a possibilidade de se obterem estruturas de tamanhos nanométricos de maneira rápida, em poucas etapas sintéticas e por meio de simples atrações físicas entre as macrocadeias, simulando as interações dos peptídeos nas proteínas. A principal característica desta técnica baseia-se na estrutura química dos polímeros sintéticos que permite o processo de autoagregação somente com interações físicas entre as macrocadeias (sem ligações covalentes). Por meio desta técnica é possível obter, com eficiência e rapidez, estruturas nanométricas que seriam de difícil obtenção por técnicas convencionais.O objetivo principal desta tese foi estudar uma rota química para a síntese de copolímeros em bloco anfifílicos e a preparação de nanopartículas sensíveis à variação de temperatura e pH, pelo método de auto-agregação. Para isso, copolímeros em bloco anfifílicos foram sintetizados utilizando como segmento hidrofóbico o Poli(hidroxibutirato-co-hidroxivalerato) (PHBHV) e como segmento hidrofílico foram utilizados a Poli(N-isopropilacrilamida) (PNIPAAm) e Poli(N-isopropilacrilamida-co-ácido acrílico) (PNIPAAmAA). Estes polímeros chamados \"inteligentes\" foram sintetizados pelo novo mecanismo de polimerização radicalar controlada, por transferência de cadeia, via fragmentação e adição reversíveis (RAFT). Essas nanopartículas termo-pH-sensíveis foram empregadas nos estudos de liberação controlada de um ativo modelo, o acetato de dexametasona, sob condições controladas de temperatura e pH. Com os resultados obtidos nesta tese foi possível identificar uma rota química de síntese de copolímeros em bloco anfifílicos sensíveis a temperatura e pH, utilizando-se de reações de acoplamento entre um polímero biodegradável, obtido de fontes renováveis, e polímeros \"inteligentes\". Foi possível demonstrar também, a viabilidade de utilização destes copolímeros anfifílicos na preparação de nanopartículas pela técnica de auto-agregação, o emprego deste sistema na encapsulação e a liberação controlada de um ativo modelo em função de variações de temperatura e pH. / Recently, the self-assembly technique provided an efficient and rapid pathway for obtaining nanometers structures in a nanometer scale using few steps of reactions and by means of simple physical attractions among macro chains, simulating the folding of peptide segments in proteins. The main characteristic of this technique is based on the chemical structure of the synthetic polymers which allow the self degradation process only with physical interactions between the macro chains (without covalent bonds). By the utilization of this technique is possible to obtain, easily and efficiently, nanometers structures, which would be difficult to be obtained by conventional techniques. The aim of this work was to study a chemical route for designing amphiphilic block copolymers and nanoparticles that exhibit thermo and pH responsive by means of self-assembly method. For this purpose, amphiphilic block copolymers were synthesized using as hydrophobic segment Poly(hydroxybutyrate-co-hydroxyvalerate) (PHBHV) and as hydrophilic segments, Poly(N-isopropylacrylamide) (PNIPAAm) and Poly(N-isopropylacrylamide-co-acrylic acid) (PNIPAAmAA). The hydrophilic polymers, called \"smart\" polymers were synthesized by a new mechanism of controlled radical polymerization, the reversible addition-fragmentation chain transfer (RAFT). These nanoparticles sensitive to temperature and pH were utilized in a drug delivery system of a model drug, the Dexametasone acetate (DexAc) under controlled environment of temperature and pH. The results allowed identifying a chemical route for the synthesis of stimuli-responsive amphiphilic block copolymers by means of coupling reactions of a biodegradable polymer obtained from renewable resources with smart polymers. It was also possible to demonstrate the possibility of utilization of these amphiphilic copolymers in the preparation of nanoparticles by self-assembly technique as well as the utilization of this system in the encapsulation and in the drug delivery of a model drug with variation of temperature and pH.

The interaction of materials with biological systems is of critical importance to a vast number of applications from medical implants, tissue engineering scaffolds, blood-contacting devices, cell-culture products, as well as many other products in industries as diverse as agriculture. This thesis describes a method for the modification of biomaterial surfaces and the generation of tissue engineering scaffolds that utilises the self assembly of poly (styrene)-block-poly (ethylene oxide) (PS-PEO) block copolymers. Block copolymers consist of alternating segments of two or more chemically distinct polymers. The salient feature of these materials is their ability to self organise into a wide range of micro-phase separated structures generating patterned surfaces that have domain sizes in the order of 10-100nm. Further, it is also possible to specifically functionalise only one segment of the block copolymer, providing a means to precisely locate specific biological signals within the 10-100nm domains of a nano-patterned surface, formed via the programmed micro-phase separation of the block copolymer system. The density and spatial location of signalling molecules can be controlled by altering several variables, such as block length, block asymmetry, as well as processing parameters, providing the potential to authentically emulate the cellular micro to nano-environment and thus greatly improving on existing biomaterial and tissue engineering technologies. This thesis achieved several aims as outlined below; Developed methods to control the self-assembly of PS-PEO block copolymers and generate nano-patterned surfaces and scaffolds with utility for biomaterials applications. PS-PEO diblock copolymers were blended with polystyrene (PS) homopolymer and spin cast, resulting in the rapid self-assembly of vertically oriented PEO cylinders in a matrix of PS. Due to the kinetically constrained phase-separation of the system, increasing addition of homopolymer is shown to reduce the diameter of the PEO domains. This outcome provides a simple method that requires the adjustment of a single variable to tune the size of vertically oriented PEO domains between 10-100nm. Polymeric scaffolds for tissue engineering were manufactured via a method that combines macro-scale temperature induced phase separation with micro-phase separation of block copolymers. The phase behaviour of these polymer-solvent systems is described, and potential mechanisms leading to this spectacular structure formation are presented. The result is highly porous scaffolds with surfaces comprised of nano-scale self-assembled block copolymer domains, representing a significant advance in currently available technologies. Characterised the properties of these unique nano-structured materials as well as their interaction with proteinaceous fluids and cells. Nano-patterned PS-PEO self-assembled surfaces showed a significant reduction in protein adsorption compared to control PS surfaces. The adhesion of NIH 3T3 fibroblast cells was shown to be significantly affected by the surface coverage of PEO nano-domains formed by copolymer self-assembly. These nano-islands, when presented at high number density (almost 1000 domains per square micron), were shown to completely prevent cellular attachment, even though small amounts of protein were able to bind to the surface. In order to understand the mechanism by which these surfaces resisted protein and cellular adsorption we utilised neutron reflection to study their solvation and swelling properties. The results indicate that the PEO domains are highly solvated in water; however, the PEO chains do not extend into the solvent but remain in their isolated domains. The data supports growing evidence that the key mechanism by which PEO prevents protein adsorption is the blocking of protein adsorption sites. Control the nano-scale presentation of cellular adhesion and other biological molecules via the self-assembly of functionalised PS-PEO block copolymers Precise control over the nano-scale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, including differentiation and apoptosis. We present the self-assembly of maleimide functionalised PS-PEO copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By controlling the phase separation of the functional PS-PEO block copolymer we alter the nano-scale (on PEO islands of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from ~ 450 to ~ 900 islands per um2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between islands of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces were utilised to immobilise poly-histidine tagged proteins and ECM fragments. The technologies developed in this thesis aim to improve on several weaknesses of existing biomaterials, in particular, directing cellular behaviour on surfaces, and within tissue engineering scaffolds, but also, on the prevention of fouling of biomaterials via non-specific protein adsorption. The application of block copolymer self-assembly for biomaterial and tissue engineering systems described in this thesis has great potential as a platform technology for the investigation of fundamental cell-surface and protein-surface interactions as well as for use in existing and emerging biomedical applications.

En la actualidad el mercado carece de prendas elásticas para uso profesional puesto que los elastanos habitualmente utilizados son inestables frente a altas temperaturas y químicos agresivos, e.g. termofijado, mantenimiento de indumentaria industrial etc. Recientemente se han venido utilizado filamentos olefínicos obtenidos a partir de copolímeros al azar de etileno-octeno (Dow XLA fibers producidos por The Dow Chemical Company). Aunque más resistentes a las temperaturas debido a la reticulación, su bajo punto de fusión, alrededor de 50ºC, limita el comportamiento elástico en tejidos pesados y tupidos para indumentaria profesional. La investigación tiene por objeto la obtención de monofilamentos elásticos a partir copolímeros de bloque de etileno-octeno (cuyo punto de fusión es más elevado) y con éstos, tejidos para indumentaria profesional que ofrezcan mayor fuerza de encogimiento a las temperaturas típicas de los pre-tratamientos, tintura y acabado (50ºC-120ºC). El objetivo es hacer que los tejidos encojan más durante las operaciones en húmedo bajo temperatura para mejorar así su elasticidad y aumentar el peso en comparación con los tejidos actuales manufacturados con los convencionales copolímeros homogéneos de etileno-octeno reticulados Los tejidos deberán garantizar las siguientes propiedades durante todo su ciclo de vida: estabilidad dimensional, relación elasticidad-crecimiento mejorado, resistencia química, resistencia térmica, durabilidad ante los lavados industriales y un mejor confort termofisiológico y sensorial. / Due to the lack of stability of elastane based materials against high temperatures and harsh chemicals, e.g. thermofixation and professional wear maintenance, elastic garments with elastane content are not yet fully commercial products. The Dow Chemical Company has recently commercialized a random ethylene-octene copolymer based fiber named Dow XLA fiber. Despite its higher temperature resistance as one of the crosslinking effects, its very low melting point, around 50ºC, establishes some limitations in elasticity such in heavy weight and high densely woven fabric cases typically found in professional wear applications. The target for the investigation is to produce block ethyleneoctene copolymer based filaments in which the melting point is supposed to be higher than in random copolymer materials. Different workwear fabrics will be manufactured with yarns containing such elastic filaments. It is supposed these fabrics to show high shrinkage forces even at dyeing and finishing temperatures, (50ºC-120ºC). The goal is to produce fabrics with superior shrinkage performance in order to achieve an excellent elastic power and the final desired fabric weight. Properties such as dimensional stability, stretch and growth ratio, temperature and chemical resistance and durability against industrial laundering should not be affected by the passage of the time. To validate this point these properties will be measured in the fabrics off loom and after the garment cycle life. These fabrics are also supposed to show better thermophysiological and sensorial properties than random copolymer fiber based textiles

Recent advancements in pharmaceutical technology based on principles of nanotechnology, polymer chemistry, and biomedical engineering have resulted in the creation of novel drug delivery systems with the potential to revolutionize current strategies in cancer chemotherapy. In oncology, realization of significant improvements in therapeutic efficacy requires minimization of drug exposure to healthy tissues and concentration of the drug within the tumor. As such, encapsulation of chemotherapeutic agents inside nanoparticles capable of enhancing tumor-targeted drug delivery is a particularly promising innovation. Yet, initial investigations into the intratumoral fate of nanomedicines have suggested that they may be heterogeneously distributed and achieve limited access to cancer cells located distant from the tumor vasculature. As such, uncovering the determinants of nanoparticle transport at the intratumoral level is critical to the development of optimized delivery vehicles capable of fully exploiting the therapeutic potential of nanomedicines. In this work, the chemotherapeutic agent, docetaxel (DTX), was incorporated into nano-sized, biocompatible PEG-b-PCL block copolymer micelles (BCMs). Encapsulation of DTX in micelles via chemical conjugation or physical entrapment resulted in a dramatic increase in drug solubility and customizable drug release rate. The use of multicellular tumor spheroids (MCTS) was established as a viable platform for assessing the efficacy and tumor tissue penetration of nanomedicines in vitro. A series of complementary assays was validated for analysis of DTX-loaded micelle (BCM+DTX) toxicity in monolayer and spheroid cultures relative to Taxotere®. Cells cultured as spheroids were less responsive to treatment relative to monolayer cultures due to mechanisms of drug resistance associated with structural and microenvironmental properties of the 3-D tissue. Computational, image-based methodologies were used to assess the spatial and temporal penetration of BCMs in spheroids and corresponding human tumor xenografts. Using this approach, the tumor penetration of micelles was found to be nanoparticle-size-, tumor tissue type- and time- dependent. Furthermore, spheroids were found to be a valuable platform for the prediction of trends in nanoparticle transport in vivo. Overall, the results reported herein serve to demonstrate important determinants of nanoparticle intratumoral transport and to establish computational in vitro and in vivo methodologies for the rational design and optimization of nanomedicines.

Recent advancements in pharmaceutical technology based on principles of nanotechnology, polymer chemistry, and biomedical engineering have resulted in the creation of novel drug delivery systems with the potential to revolutionize current strategies in cancer chemotherapy. In oncology, realization of significant improvements in therapeutic efficacy requires minimization of drug exposure to healthy tissues and concentration of the drug within the tumor. As such, encapsulation of chemotherapeutic agents inside nanoparticles capable of enhancing tumor-targeted drug delivery is a particularly promising innovation. Yet, initial investigations into the intratumoral fate of nanomedicines have suggested that they may be heterogeneously distributed and achieve limited access to cancer cells located distant from the tumor vasculature. As such, uncovering the determinants of nanoparticle transport at the intratumoral level is critical to the development of optimized delivery vehicles capable of fully exploiting the therapeutic potential of nanomedicines. In this work, the chemotherapeutic agent, docetaxel (DTX), was incorporated into nano-sized, biocompatible PEG-b-PCL block copolymer micelles (BCMs). Encapsulation of DTX in micelles via chemical conjugation or physical entrapment resulted in a dramatic increase in drug solubility and customizable drug release rate. The use of multicellular tumor spheroids (MCTS) was established as a viable platform for assessing the efficacy and tumor tissue penetration of nanomedicines in vitro. A series of complementary assays was validated for analysis of DTX-loaded micelle (BCM+DTX) toxicity in monolayer and spheroid cultures relative to Taxotere®. Cells cultured as spheroids were less responsive to treatment relative to monolayer cultures due to mechanisms of drug resistance associated with structural and microenvironmental properties of the 3-D tissue. Computational, image-based methodologies were used to assess the spatial and temporal penetration of BCMs in spheroids and corresponding human tumor xenografts. Using this approach, the tumor penetration of micelles was found to be nanoparticle-size-, tumor tissue type- and time- dependent. Furthermore, spheroids were found to be a valuable platform for the prediction of trends in nanoparticle transport in vivo. Overall, the results reported herein serve to demonstrate important determinants of nanoparticle intratumoral transport and to establish computational in vitro and in vivo methodologies for the rational design and optimization of nanomedicines.

The interaction of materials with biological systems is of critical importance to a vast number of applications from medical implants, tissue engineering scaffolds, blood-contacting devices, cell-culture products, as well as many other products in industries as diverse as agriculture. This thesis describes a method for the modification of biomaterial surfaces and the generation of tissue engineering scaffolds that utilises the self assembly of poly (styrene)-block-poly (ethylene oxide) (PS-PEO) block copolymers. Block copolymers consist of alternating segments of two or more chemically distinct polymers. The salient feature of these materials is their ability to self organise into a wide range of micro-phase separated structures generating patterned surfaces that have domain sizes in the order of 10-100nm. Further, it is also possible to specifically functionalise only one segment of the block copolymer, providing a means to precisely locate specific biological signals within the 10-100nm domains of a nano-patterned surface, formed via the programmed micro-phase separation of the block copolymer system. The density and spatial location of signalling molecules can be controlled by altering several variables, such as block length, block asymmetry, as well as processing parameters, providing the potential to authentically emulate the cellular micro to nano-environment and thus greatly improving on existing biomaterial and tissue engineering technologies. This thesis achieved several aims as outlined below; Developed methods to control the self-assembly of PS-PEO block copolymers and generate nano-patterned surfaces and scaffolds with utility for biomaterials applications. PS-PEO diblock copolymers were blended with polystyrene (PS) homopolymer and spin cast, resulting in the rapid self-assembly of vertically oriented PEO cylinders in a matrix of PS. Due to the kinetically constrained phase-separation of the system, increasing addition of homopolymer is shown to reduce the diameter of the PEO domains. This outcome provides a simple method that requires the adjustment of a single variable to tune the size of vertically oriented PEO domains between 10-100nm. Polymeric scaffolds for tissue engineering were manufactured via a method that combines macro-scale temperature induced phase separation with micro-phase separation of block copolymers. The phase behaviour of these polymer-solvent systems is described, and potential mechanisms leading to this spectacular structure formation are presented. The result is highly porous scaffolds with surfaces comprised of nano-scale self-assembled block copolymer domains, representing a significant advance in currently available technologies. Characterised the properties of these unique nano-structured materials as well as their interaction with proteinaceous fluids and cells. Nano-patterned PS-PEO self-assembled surfaces showed a significant reduction in protein adsorption compared to control PS surfaces. The adhesion of NIH 3T3 fibroblast cells was shown to be significantly affected by the surface coverage of PEO nano-domains formed by copolymer self-assembly. These nano-islands, when presented at high number density (almost 1000 domains per square micron), were shown to completely prevent cellular attachment, even though small amounts of protein were able to bind to the surface. In order to understand the mechanism by which these surfaces resisted protein and cellular adsorption we utilised neutron reflection to study their solvation and swelling properties. The results indicate that the PEO domains are highly solvated in water; however, the PEO chains do not extend into the solvent but remain in their isolated domains. The data supports growing evidence that the key mechanism by which PEO prevents protein adsorption is the blocking of protein adsorption sites. Control the nano-scale presentation of cellular adhesion and other biological molecules via the self-assembly of functionalised PS-PEO block copolymers Precise control over the nano-scale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, including differentiation and apoptosis. We present the self-assembly of maleimide functionalised PS-PEO copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By controlling the phase separation of the functional PS-PEO block copolymer we alter the nano-scale (on PEO islands of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from ~ 450 to ~ 900 islands per um2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between islands of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces were utilised to immobilise poly-histidine tagged proteins and ECM fragments. The technologies developed in this thesis aim to improve on several weaknesses of existing biomaterials, in particular, directing cellular behaviour on surfaces, and within tissue engineering scaffolds, but also, on the prevention of fouling of biomaterials via non-specific protein adsorption. The application of block copolymer self-assembly for biomaterial and tissue engineering systems described in this thesis has great potential as a platform technology for the investigation of fundamental cell-surface and protein-surface interactions as well as for use in existing and emerging biomedical applications.

The interaction of materials with biological systems is of critical importance to a vast number of applications from medical implants, tissue engineering scaffolds, blood-contacting devices, cell-culture products, as well as many other products in industries as diverse as agriculture. This thesis describes a method for the modification of biomaterial surfaces and the generation of tissue engineering scaffolds that utilises the self assembly of poly (styrene)-block-poly (ethylene oxide) (PS-PEO) block copolymers. Block copolymers consist of alternating segments of two or more chemically distinct polymers. The salient feature of these materials is their ability to self organise into a wide range of micro-phase separated structures generating patterned surfaces that have domain sizes in the order of 10-100nm. Further, it is also possible to specifically functionalise only one segment of the block copolymer, providing a means to precisely locate specific biological signals within the 10-100nm domains of a nano-patterned surface, formed via the programmed micro-phase separation of the block copolymer system. The density and spatial location of signalling molecules can be controlled by altering several variables, such as block length, block asymmetry, as well as processing parameters, providing the potential to authentically emulate the cellular micro to nano-environment and thus greatly improving on existing biomaterial and tissue engineering technologies. This thesis achieved several aims as outlined below; Developed methods to control the self-assembly of PS-PEO block copolymers and generate nano-patterned surfaces and scaffolds with utility for biomaterials applications. PS-PEO diblock copolymers were blended with polystyrene (PS) homopolymer and spin cast, resulting in the rapid self-assembly of vertically oriented PEO cylinders in a matrix of PS. Due to the kinetically constrained phase-separation of the system, increasing addition of homopolymer is shown to reduce the diameter of the PEO domains. This outcome provides a simple method that requires the adjustment of a single variable to tune the size of vertically oriented PEO domains between 10-100nm. Polymeric scaffolds for tissue engineering were manufactured via a method that combines macro-scale temperature induced phase separation with micro-phase separation of block copolymers. The phase behaviour of these polymer-solvent systems is described, and potential mechanisms leading to this spectacular structure formation are presented. The result is highly porous scaffolds with surfaces comprised of nano-scale self-assembled block copolymer domains, representing a significant advance in currently available technologies. Characterised the properties of these unique nano-structured materials as well as their interaction with proteinaceous fluids and cells. Nano-patterned PS-PEO self-assembled surfaces showed a significant reduction in protein adsorption compared to control PS surfaces. The adhesion of NIH 3T3 fibroblast cells was shown to be significantly affected by the surface coverage of PEO nano-domains formed by copolymer self-assembly. These nano-islands, when presented at high number density (almost 1000 domains per square micron), were shown to completely prevent cellular attachment, even though small amounts of protein were able to bind to the surface. In order to understand the mechanism by which these surfaces resisted protein and cellular adsorption we utilised neutron reflection to study their solvation and swelling properties. The results indicate that the PEO domains are highly solvated in water; however, the PEO chains do not extend into the solvent but remain in their isolated domains. The data supports growing evidence that the key mechanism by which PEO prevents protein adsorption is the blocking of protein adsorption sites. Control the nano-scale presentation of cellular adhesion and other biological molecules via the self-assembly of functionalised PS-PEO block copolymers Precise control over the nano-scale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, including differentiation and apoptosis. We present the self-assembly of maleimide functionalised PS-PEO copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By controlling the phase separation of the functional PS-PEO block copolymer we alter the nano-scale (on PEO islands of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from ~ 450 to ~ 900 islands per um2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between islands of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces were utilised to immobilise poly-histidine tagged proteins and ECM fragments. The technologies developed in this thesis aim to improve on several weaknesses of existing biomaterials, in particular, directing cellular behaviour on surfaces, and within tissue engineering scaffolds, but also, on the prevention of fouling of biomaterials via non-specific protein adsorption. The application of block copolymer self-assembly for biomaterial and tissue engineering systems described in this thesis has great potential as a platform technology for the investigation of fundamental cell-surface and protein-surface interactions as well as for use in existing and emerging biomedical applications.

The interaction of materials with biological systems is of critical importance to a vast number of applications from medical implants, tissue engineering scaffolds, blood-contacting devices, cell-culture products, as well as many other products in industries as diverse as agriculture. This thesis describes a method for the modification of biomaterial surfaces and the generation of tissue engineering scaffolds that utilises the self assembly of poly (styrene)-block-poly (ethylene oxide) (PS-PEO) block copolymers. Block copolymers consist of alternating segments of two or more chemically distinct polymers. The salient feature of these materials is their ability to self organise into a wide range of micro-phase separated structures generating patterned surfaces that have domain sizes in the order of 10-100nm. Further, it is also possible to specifically functionalise only one segment of the block copolymer, providing a means to precisely locate specific biological signals within the 10-100nm domains of a nano-patterned surface, formed via the programmed micro-phase separation of the block copolymer system. The density and spatial location of signalling molecules can be controlled by altering several variables, such as block length, block asymmetry, as well as processing parameters, providing the potential to authentically emulate the cellular micro to nano-environment and thus greatly improving on existing biomaterial and tissue engineering technologies. This thesis achieved several aims as outlined below; Developed methods to control the self-assembly of PS-PEO block copolymers and generate nano-patterned surfaces and scaffolds with utility for biomaterials applications. PS-PEO diblock copolymers were blended with polystyrene (PS) homopolymer and spin cast, resulting in the rapid self-assembly of vertically oriented PEO cylinders in a matrix of PS. Due to the kinetically constrained phase-separation of the system, increasing addition of homopolymer is shown to reduce the diameter of the PEO domains. This outcome provides a simple method that requires the adjustment of a single variable to tune the size of vertically oriented PEO domains between 10-100nm. Polymeric scaffolds for tissue engineering were manufactured via a method that combines macro-scale temperature induced phase separation with micro-phase separation of block copolymers. The phase behaviour of these polymer-solvent systems is described, and potential mechanisms leading to this spectacular structure formation are presented. The result is highly porous scaffolds with surfaces comprised of nano-scale self-assembled block copolymer domains, representing a significant advance in currently available technologies. Characterised the properties of these unique nano-structured materials as well as their interaction with proteinaceous fluids and cells. Nano-patterned PS-PEO self-assembled surfaces showed a significant reduction in protein adsorption compared to control PS surfaces. The adhesion of NIH 3T3 fibroblast cells was shown to be significantly affected by the surface coverage of PEO nano-domains formed by copolymer self-assembly. These nano-islands, when presented at high number density (almost 1000 domains per square micron), were shown to completely prevent cellular attachment, even though small amounts of protein were able to bind to the surface. In order to understand the mechanism by which these surfaces resisted protein and cellular adsorption we utilised neutron reflection to study their solvation and swelling properties. The results indicate that the PEO domains are highly solvated in water; however, the PEO chains do not extend into the solvent but remain in their isolated domains. The data supports growing evidence that the key mechanism by which PEO prevents protein adsorption is the blocking of protein adsorption sites. Control the nano-scale presentation of cellular adhesion and other biological molecules via the self-assembly of functionalised PS-PEO block copolymers Precise control over the nano-scale presentation of adhesion molecules and other biological factors represents a new frontier for biomaterials science. Recently, the control of integrin spacing and cellular shape has been shown to affect fundamental biological processes, including differentiation and apoptosis. We present the self-assembly of maleimide functionalised PS-PEO copolymers as a simple, yet highly precise method for controlling the position of cellular adhesion molecules. By controlling the phase separation of the functional PS-PEO block copolymer we alter the nano-scale (on PEO islands of 8-14 nm in size) presentation of the adhesion peptide, GRGDS, decreasing lateral spacing from 62 nm to 44 nm and increasing the number density from ~ 450 to ~ 900 islands per um2. The results indicate that the spreading of NIH-3T3 fibroblasts increases as the spacing between islands of RGD binding peptides decreases. Further, the same functional PS-PEO surfaces were utilised to immobilise poly-histidine tagged proteins and ECM fragments. The technologies developed in this thesis aim to improve on several weaknesses of existing biomaterials, in particular, directing cellular behaviour on surfaces, and within tissue engineering scaffolds, but also, on the prevention of fouling of biomaterials via non-specific protein adsorption. The application of block copolymer self-assembly for biomaterial and tissue engineering systems described in this thesis has great potential as a platform technology for the investigation of fundamental cell-surface and protein-surface interactions as well as for use in existing and emerging biomedical applications.

Les nanoparticules magnétiques sont de nos jours largement exploitées dans le domaine de la recherche pour le biomédical, pour des applications aussi variées que le diagnostic, la thérapeutique ou plus récemment la théranostique. Les nombreuses méthodes de fabrication mises au point à ce jour permettent l’obtention d’une large gamme de nanoparticules en termes de taille, forme, matériaux et donc propriétés magnétiques. Le procédé de fabrication idéal est celui qui permet la fabrication simple, peu coûteuse et à grande échelle de nanoparticules parfaitement monodisperses. En ce sens, le premier volet de la thèse sera consacré à l’étude d’un nouveau procédé de fabrication basé sur la combinaison d’une approche « top-down » et « bottom-up », qui permet d’obtenir des nanoparticules de dispersion en taille très étroite. L’idée est d’exploiter les propriétés d’auto-organisation d’un copolymère dibloc, dont l’une des deux phases peut s’organiser en cylindres verticaux dans la matrice de l’autre phase sous certaines conditions. La gravure sélective des cylindres mène à l’obtention d’un masque de trous dans une matrice de polymère. On peut ensuite déposer le matériau magnétique, puis graver la matrice de polymère pour révéler les nanoparticules attachées au substrat. Si ce procédé est mené sur une couche sacrificielle, les particules peuvent être consécutivement mises en suspension. Les caractéristiques structurales et magnétiques de ces particules obtenues par auto-organisation du copolymère PS-PMMA seront étudiées, montrant que bien que ce procédé de fabrication soit encore à améliorer, il présente des avantages non négligeables en étant versatile, simple à mettre en œuvre et en permettant l’obtention de nanoparticules monodisperses et superparamagnétiques.Dans une seconde partie, un autre domaine biomédical sera abordé : le traitement du cancer. Une méthode nouvelle et alternative aux techniques d’hyperthermie ou de délivrance ciblée de médicaments avait été initiée par l’Argonne National Laboratory en 2010 et reprise à Spintec en 2011 : l’idée est de réactiver l’apoptose (ou mort programmée) de cellules cancéreuses par vibrations magnéto-mécaniques de microparticules magnétiques attachées à leur membrane. Il avait été démontré qu’avec des champs extérieurs aussi faibles que 30mT à 20Hz, des disques de permalloy en configuration magnétique vortex induisent l’apoptose de façon significative. Dans ce contexte et dans l’optique de pouvoir utiliser cette méthode pour des tests cliniques, des microparticules de magnétite, matériau biocompatible, ont été fabriquées par lithographie optique via le même procédé que les disques de permalloy. Leurs propriétés structurales, magnétiques et leur comportement en suspension sont comparés, ainsi que leurs effets sur les cellules in vitro via l’application d’un champ magnétique extérieur. A ce jour, les particules de permalloy sont supérieures en termes d’efficacité sur le déclenchement de l’apoptose des cellules cancéreuses. Certains paramètres du protocole tels que l’amplitude du champ doivent être optimisés pour les particules de magnétite, bien que les premiers effets observés soient encourageants pour la suite. / Magnetic nanoparticles are now used in a wide range of applications such as diagnostic, therapeutics or more recently theranostics. The numerous and diverse fabrication processes allow the fabrication of a wide range of nanoparticles in terms of size, shape, material and magnetic properties. An ideal fabrication process would allow the simple and cheap fabrication of a great quantity of monodisperse nanoparticles. In this objective, the first part of this work will be focused on a new and original fabrication process based on the combination of a “top-down” and “bottom-up” approach. The idea relies on the special auto organization properties of a diblock copolymer: one of the two phases has the ability to self organize into vertical cylinders in the matrix of the other polymer, provided that the annealing conditions are favourable. The selective etching of the cylinders leads to a mask of holes in a polymer matrix. Then, the deposit of a magnetic material and the etching of the polymer matrix leads to the formation of a hexagonal network of nanoparticles attached to the substrate. If the substrate is composed of a sacrificial layer, the nanoparticles can be released in a solution. The structural and magnetic properties of theses nanoparticles fabricated via a PS-PMMA template will be studied. Their characterization will show that the process is still to be optimized but allows already to obtain monodisperse superparamagnetic nanoparticles.A second part focuses on another biomedical applications of magnetic particles: the cancer treatment. A new technique, which is an alternative to the existing methods such as hyperthermia or drug delivery, was first proposed by the Argonne National Laboratory (2010) and taken over at Spintec (2011). The idea is to reactivate the apoptosis (programmed cell death) of cancer cells via the magneto mechanical vibrations of magnetic microparticles attached to their membranes. It was proved that weak external magnetic fields (30mT at 20Hz) applied on permalloy disks in a vortex configuration lead to a significant increase of the apoptotic rate of cancer cells. In the objective of making this method possible for clinical applications, biocompatible magnetite microparticles were fabricated via the same fabrication process than the permalloy disks (optic lithography). Their structural and magnetic properties are compared, as well as their behavior in a suspension and their lethal effect on cancer cells via the application of an external magnetic field. For now, the permalloy microdisks provide better results than the magnetite particles. Some parameters of the experimental set up have to be optimized for the magnetite particles, such as the amplitude of the applied magnetic field. However, the first effects observed with the magnetite particles are quite promising.

Orientador: Prof. Dr. Danilo Justino Carastan / Dissertação (mestrado) - Universidade Federal do ABC, Programa de Pós-Graduação em Nanociências e Materiais Avançados, 2015. / Copolímeros em bloco são materiais conhecidos pela capacidade de formar diferentes estruturas ordenadas na escala nanométrica. A adição de nanopartículas a estes copolímeros possibilita formar nanocompósitos com propriedades e morfologias interessantes, dependendo da interação entre a nanopartícula e os domínios dos blocos, além das condições de processamento. É possível orientar copolímeros em bloco resultando em estruturas anisotrópicas, através de fluxos de cisalhamento e elongacional. Neste trabalho, foram obtidos nanocompósitos do copolímero em bloco poliestireno-b-poli(etileno-co-butileno)-b-poliestireno (SEBS), com morfologia hexagonal cilíndrica, com adição de diferentes nanopartículas, através da técnica de processamento de extrusão de filme tubular. A principal finalidade é estudar a orientação morfológica, uniaxial e biaxial, das amostras em função da razão de inflamento, durante o inflamento do filme tubular, e a influência das diferentes morfologias das nanopartículas adicionadas na interação com os domínios do SEBS. A alteração da estrutura morfológica é acompanhada por análises de espalhamento de raios x a baixos ângulos (SAXS) e a orientação morfológica é quantificada pelo modelo matemático do parâmetro de ordem (F). Ensaios reológicos no fluxo elongacional são realizados a fim de correlacionar o comportamento da estrutura morfológica durante o inflamento do filme tubular, que possui deformação predominantemente do tipo elongacional biaxial, e compreender seu mecanismo de deformação. Os nanocompósitos com estruturas morfológicas orientadas, uniaxial e biaxialmente, são submetidos a ensaios de tração para avaliar as propriedades mecânicas em diferentes direções de ensaio. / Block copolymers are materials known for the ability to form different ordered structures at the nanoscale. The addition of nanoparticles in these copolymers allows to form nanocomposites with interesting properties and morphologies, depending on the interaction between the nanoparticle and the domains of the blocks, in addition to the processing conditions. It is possible to align block copolymers resulting in anisotropic structures through shear and elongational flow. In this work, nanocomposites were obtained from the combination of a block copolymer polystyrene-b-poly(ethylene-co-butylene)-b-polystyrene (SEBS) with hexagonal cylindrical morphology, and different nanoparticles, through the processing technique of blown film extrusion. The main purpose is to study the morphological orientation, uniaxial and biaxial, of the samples as a function of blown-up ratio during the inflation of the tubular film, and the influence of different nanoparticle morphologies on the interaction with the SEBS domains. The change in the morphological structure is evaluated by small angle x-ray scattering (SAXS) analysis and morphological orientation is quantified by the mathematical model of the order parameter (F). Rheological elongational flows are performed in order to correlate the behavior of the morphological structure during the inflation of the tubular film, which has predominantly elongational deformation of the biaxial type, and understand their deformation mechanism. Nanocomposites with oriented morphological structures, uniaxial and biaxial, are subjected to tensile tests to evaluate the mechanical properties in different test directions.