Rozprawy doktorskie na temat „Biodegradable polymeric nanoparticles”

Multiple hurdles, such as drug solubility, stability, and physical barriers in the body, hinder bioavailability of many promising therapeutics. Polymeric nanocarriers can encapsulate the therapeutics to protect non-target areas from side effects but also protect the drug from premature degradation for increased circulation and bioavailability. To capitalize on these advantages, the polymer nanoparticle must be properly engineered for increased control in size distribution, therapeutic encapsulation, colloidal stability, and release kinetics. However, each application requires a specific set of characteristics and properties. Being able to tailor these by manipulation of different design parameters is essential to optimize nanoparticles for the application of interest. This study of nanoparticle fabrication and characterization takes us a step closer to building effective delivery systems tailored for specific treatments. Poly(ethylene oxide)-b-poly(D,L-lactic acid) (PEO-b-PDLLA) based nanoparticles were produced to range from 100-200 nm in size. They were fluorescently labeled with a hydrophobic dye 6-13 bis(triisopropylsilylethynyl) pentacene (TIPS pentacene) at an optimal loading of 0.5 wt% with respect to the core. Surfaces were successfully coated with streptavidin to be readily functionalized with various biotinylated compounds such as PD-L1 antibodies or A488 fluorophore. Using the same PEO-b-PDLLA, iron oxide and a conjugated polymer poly(2- methoxy-5-(2-ethylhexyloxy)-1,4-phenylenevinylene) (MEH-PPV) were co-encapsulated to form fluorescently labeled magnetic particles. Using poly(lactic-co-glycolic acid), CRISPR-Cas9 plasmids were encapsulated at 1.6 wt% and most of the payload released within the first 24 hours. The incorporated plasmids were intact enough to have mammalian macrophages successfully express the bacterial protein Cas9. Using similar PLGA based particles, the surface was functionalized with streptavidin and bound to the surface of bacteria as an active carrier for increased penetration of solid tumors averaging ~23 particles per bacterium. PEO-b-PLGA based particles were used in conjunction with a hydrophobic salt former to encapsulate a peptide designed to reduce platelet binding to cancer cells and mitigate extravasation. The peptide encapsulated was increased from < 2 wt% without salt former to 8.5 wt% with the used of hexadecyl phosphonic acid. Although the applications across these projects can be broad, the fundamentals and important design parameters considered contribute to the overarching field of effective carriers for drug delivery.
Ph. D.

Thesis (Ph.D.)--Biomedical Engineering, Georgia Institute of Technology, 2008.
Committee Chair: Dr. Niren Murthy; Committee Member: Dr. Carson Meredith; Committee Member: Dr. Julia Babensee; Committee Member: Dr. Mark Prausnitz; Committee Member: Dr. Ravi Bellamkonda.

Rheumatoid arthritis (RA) is an autoimmune disease affecting joints due to the persistent inflammation of the synovial tissue. It affects 1% of the worldwide population with high socioeconomic costs. Patients usually require lifetime treatments in order to prevent late stage of the disease, which leads to a condition of disability and pain. Despite the introduction of biological drugs, Methotrexate (MTX) is still the gold standard. However, it shows some weakness such as inefficacy or adverse events after long term usage. For this reason, there is the need to develop drugs with an improved safety profile and higher therapeutic efficacy. Moreover, it is of primary importance to develop diagnostics with enhanced sensitivity and tissue specificity. In order to meet these needs, the purpose of this work is to develop a nanotechnological agent capable to delivery its content, such as drugs or diagnostic tracers, specifically into pathological joints, avoiding healthy tissues. These aims have been achieved exploiting peculiar features of biodegradable nanoparticles (BNPs), such as improved pharmacokinetic and bioavailability, associated to a peptide specific for the inflamed synovia. Recently, the heterogeneity of the molecules present in the endothelium has allowed to develop peptides capable to target vessels of specific tissues. In this project, a cyclic peptide able to target the microvasculature of the inflamed synovia has been exploited to drive BNPs only into inflamed joints. BNPs have been characterized for their physicochemical features, polymers’ toxicity and drug release. Targeted polymeric nanoparticles (tBNPs) demonstrated, both in vitro and in vivo, to preferentially bind inflamed synovial tissue, and their accumulation depends from the degree of inflammation. Targeted BNPs loaded with MTX showed higher efficacy compared to free MTX in two different animal models of RA. In addition, reduced dose of tBNPs-MTX demonstrated to maintain its efficacy, showing fewer side effects compared with systemic administration of free MTX. This new therapeutic approach provided a new mechanism of action from approved therapy and suggest potential application in non-responding patients. All these evidences highlight the potential use of tBNPs as biocompatible and adaptable tool for the diagnosis and the treatment of RA showing important efficacy, reduced side effects and with the potential to enhance the sensitivity of several imaging technologies.

Tumor Associated Macrophages (TAMs) are involved in cancer proliferation, thus strategies to deplete them represent promising tools for chemotherapy. Pharmacological agents with multiple activities such as curcumin and RNA interference have been proposed; however their employment in therapeutics has been limited because of low systemic bioavailability. Accordingly, this thesis described as an innovative therapeutic approach for cancer treatment the development of polymeric nanoparticles (NPs) able to (i) increase pharmacokinetics properties of biomacromolecules and poor water soluble drugs, and (ii) guarantee TAMs specific targeting. The safe and versatile polymer Poly(Lactic-co-Glycolic)Acid (PLGA) has been used for the synthesis of NPs by both single (OW) and double (WOW) emulsion-solvent evaporation techniques. Different synthetic parameters have been taken into consideration, with particular focus on the surfactant role. As alternative to the commonly used Poly Vinyl Alcohol (PVA), a newly synthetized polymer (amino-PVA) and Calcium Sterate (CSt) have been investigated for their ability to modulate surface charge and biocompatibility. NPs with solid or core-shell structures, whose size was tailored between 200 and 300 nm, were obtained and a thorough characterization has been performed, with the help of innovative techniques such as single particle optical extinction and scattering (SPES) method. Both amino-PVA and CSt stabilized NPs were found to be able to load curcumin and biomacromolecules, either alone or in combination. Strategies for surface decoration with the employment of D-mannose as specific molecule to guarantee TAMs recognition were proposed. Finally, cytocompatibility of the amino-PVA and CSt stabilized NPs have been assessed.

Tissue engineering is a promising alternative strategy to produce artificial bone substitutes
however, the control of the cell organization and cell behavior to create fully functional 3-D constructs has not yet been achieved. To overcome these, activities have been concentrated on the development of multi-functional tissue engineering scaffolds capable of delivering the required bioactive agents to initiate and control cellular activities. The aim of this study was to prepare tissue engineered constructs composed of polymeric scaffolds seeded with mesenchymal stem cells (MSCs) carrying a nanoparticulate growth factor delivery system that would sequentially deliver the growth factors in order to mimic the natural bone healing process. To achieve this, BMP-2 and BMP-7, the osteogenic growth factors, were encapsulated in different polymeric nanocapsules (poly(lactic acid-co-glycolic acid) (PLGA) and poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV)) with different properties (degradation rates, crystallinity) and, therefore, different release rates to achieve the early release of BMP-2 followed by the release of BMP-7, as it is in nature. Initially, these nanoparticulate delivery systems were characterized and then the effect of single, simultaneous and sequential delivery of BMP-2 and BMP-7 from these delivery systems was studied in vitro using rat bone marrow MSCs. The effect of using these two growth factors in a sequential manner by mimicking their natural bioavailability timing was shown with maximized osteogenic activity results. BMP-2 loaded PLGA nanocapsules were subcutaneously implanted into Wistar rats and according to initial results, their biocompatibility as well as the positive effect of BMP-2 release on the formation of osteoclast-like cells was shown. To complete the construction of the bioactive scaffold, this nanoparticulate sequential delivery system was incorporated into two different types of polymeric systems
natural (chitosan) and synthetic (poly(&
#949
-caprolactone) (PCL)). 3-D fibrous scaffolds were produced using these materials by wet spinning and 3-D plotting. Incorporation of nanocapsules into 3-D chitosan scaffolds was studied by two different methods: incorporation within and onto chitosan fibers. Incorporation into 3-D PCL scaffolds was achieved by coating the nanocapsules onto the fibers of the scaffolds in an alginate layer. With both scaffold systems, incorporation of nanocapsule populations capable of delivering BMP-2 and BMP-7 in single, simultaneous and sequential fashion was achieved. As with free nanocapsules, the positive effect of sequential delivery on the osteogenic differentiation of MSCs was shown with both scaffold systems, creating multi-functional scaffolds capable of inducing bone healing.

This research is directed toward understanding the effect of nanoparticle attributes and polymer morphology on the properties of the nanocomposites with analogous nanoparticle chemistry. In order to develop this understanding, polymer nanocomposites containing calcium phosphate nanoparticles of different specific surface areas and shapes were fabricated and characterized through thermal and thermomechanical techniques. Nanoparticles were synthesized using reverse microemulsion technique. For nanocomposites with different surface area particles, the mobility of amorphous polymer chains was restricted significantly by the presence of particles with an interphase network morphology at higher loadings. Composites fabricated with different crystallinity matrices showed that the dispersion characteristics and reinforcement behavior of nanoparticles were governed by the amount of amorphous polymer fraction available. The study conducted on the effect of nanoparticle shape with near-spherical and nanofiber nanoparticles illustrated that the crystallization kinetics and the final microstructure of the composites was a function of shape of the nanoparticles. The results of this research indicate that nanoparticle geometry and matrix morphology are important parameters to be considered in designing and characterizing the structure-property relationship in polymer nanocomposites.

Doctor of Philosophy
Department of Grain Science and Industry
X. Susan Sun
Biobased polymers derived from renewable resources are increasingly important due to acute concerns about the environmental issues and limited petroleum resources. Poly(lactic acid) (PLA) is such a polymer that has shown great potential to produce biodegradable plastics. However, low glass transition temperature (Tg), low thermal stability, slow biodegradation rate, and high cost limit its broad applications. This dissertation seeks to overcome these limitations by reinforcing PLA with inorganic nanoparticles and low-cost agricultural residues. We first synthesized PLA nanocomposites by in situ melt polycondensation of L-lactic acid and surface-hydroxylized nanoparticles (MgO nanocrystals and TiO2 nanowires) and investigated the structure-property relationships. PLA grafted nanoparticles (PLA-g-MgO, PLA-g-TiO2) were isolated from the bulk nanocomposites via repeated dispersion/centrifugation processes. The covalent grafting of PLA chains onto nanoparticle surface was confirmed by Fourier transform infrared spectroscopy and thermalgravimetric analysis (TGA). Transmission electron microscopy and differential scanning calorimetry (DSC) results also sustained the presence of the third phase. Morphological images showed uniform dispersion of nanoparticles in the PLA matrix and demonstrated a strong interfacial interaction between them. Calculation based on TGA revealed that more than 42.5% PLA was successfully grafted into PLA-g-MgO and more than 30% was grafted into PLA-g-TiO2. Those grafted PLA chains exhibited significantly increased thermal stability. The Tg of PLA-g-TiO2 was improved by 7 °C compared with that of pure PLA. We also reinforced PLA with low-value agricultural residues, including wood flour (WF), soy flour (SF), and distillers dried grains with solubles (DDGS) by thermal blending. Tensile measurements and morphological images indicated that methylene diphenyl diisocyanate (MDI) was an effective coupling agent for PLA/WF and PLA/DDGS systems. MDI compatibilized PLA/WF and PLA/DDGS composites showed comparable tensile strength and elongation at break as pure PLA, with obviously increased Young’s modulus. Increased crystallinity was observed for PLA composites with SF and DDGS. Such PLA composites have similar or superior properties compared with pure PLA, especially at a lower cost and higher biodegradation rate than pure PLA. The results from this study are promising. These novel PLA thermoplastic composites with enhanced properties have potential for many applications, such as packaging materials, textiles, appliance components, autoparts, and medical implants.

The aims of this study were to develop polymeric NP formulations for triamcinolone acetonide (TA) delivery, from biodegradable and biocompatible hydrophobic polymers, which provide sustained release, prolonged stability and low toxicity, and to assess the toxicity of TA NPs (TA-NPs) compared to TA alone upon BALB/c 3T3 and ARPE 19 cell culture models.The study involved investigation of three different types of polymers: poly(D,L,lactide) (PDLLA), poly(D,L,lactide-co-glycolide)(PLGA) and methoxypolyethyleneglycol poly(D,L,lactide-co-glycolide)(mPEG PLGA). Two different methods were studied in the TA-NPs preparation: spontaneous emulsification solvent diffusion and emulsification solvent evaporation methods.The results show that emulsification-solvent evaporation method was superior to spontaneous emulsification solvent diffusion in terms of yield, loading and entrapment efficiency. TA-NPs synthesised of mPEG PLGA exhibited the smallest particle size, highest efficiency and fastest release of TA, whereas PDLLA produced large TA-NPs with the slowest TA release. The toxicity study revealed that BALB/c 3T3 was more sensitive than ARPE 19 and was concentration dependent in response to 24 hour exposure of either TA or TA-NPs, while ARPE 19 appeared to be less sensitive to the exposure. All NPs were less toxic than TA in all concentrations, in both cell models.

Hydrogels are popular materials for biological applications since they exhibit properties like that of natural soft tissue and have tunable properties. Biodegradable hydrogels provide an added advantage in that they degrade in an aqueous environment thereby avoiding the need for removal after the useful lifetime. In this work, we investigated poly(β-amino ester) (PBAE) biodegradable hydrogel systems. To begin, the factors affecting the macromer synthesis procedure were studied to optimize the reproducibility of the resulting hydrogels made and create new methods of tuning the properties. Hydrogel behavior was then tuned by altering the hydrophilic/hydrophobic balance of the chemicals used in the synthesis to develop systems with linear and two-phase degradation profiles. The goal of the research was to better understand methods of controlling hydrogel properties to develop systems for several biomedical applications. Several systems with a range of properties were synthesized, and their in vitro behavior was characterized (degradation, mechanical properties, cellular response, etc.). From these studies, materials were chosen to serve as porogen materials and an outer matrix material to create a composite scaffold for tissue engineering. In most cases, a porous three dimensional scaffold is ideal for cellular growth and infiltration. In this work, a composite with a slow degrading outer matrix PBAE with fast degrading PBAE microparticles was created. First, a procedure for developing porogen particles of controlled size from a fast-degrading hydrogel material was developed. Porogen particles were then entrapped in the outer hydrogel matrix during polymerization. The resulting composite systems were degraded and the viability of these systems as tissue engineering scaffolds was studied. In a second area of work, two polymer systems, one PBAE hydrogel and one sol-gel material were altered through the addition of iron oxide nanoparticles to create materials with remote controlled properties. Iron oxide nanoparticles have the ability to heat in an alternating magnetic field due to the relaxation processes. The incorporation of these nanoscale heating sources into thermosensitive polymer systems allowed remote actuation of the physical properties. These materials would be ideal for use in applications where the system can be changed externally such as in remote controlled drug delivery.

Le traitement des infections intracellulaires est compliqué par la capacité des bactéries à «se cacher» à l’intérieur des cellules de l’hôte, en particulier celles du système immunitaire, entravant ainsi l’action de nombreux agents antimicrobiens. La diffusion croissante de souches résistantes est très inquiétante. Dans ce cadre, les nanoparticules (NPs) constituent une stratégie prometteuse pour administrer de manière optimisée des agents antimicrobiens.Ce travail de thèse, réalisé dans le cadre du projet européen ITN Cyclon Hit, visait à développer et caractériser des NPs biodégradables et biocompatibles chargées en antibiotiques, composés d’acide polylactique (PLA), d’acide poly (lactique-co-glycolique) (PLGA) et de polycaprolactone (PCL) ou de cyclodextrines polymérisées (pCD).Les deux premiers chapitres sont consacrés aux verrous technologiques liés à l'encapsulation de certains médicaments puissants dans les NPs polymériques. Tout d'abord, ces vecteurs ont été utilisés pour la délivrance simultanée d'une combinaison de molécules actives récemment découverte, l'éthionamide (ETH) et son Booster, pour le traitement de la tuberculose. Deuxièmement, ils ont été employés pour relever les défis liés à l'incorporation d'une quinolone de première génération, l'acide pipémidique (PIP), dans le but d'optimiser sa distribution intracellulaire dans des infections telles que la salmonellose.La co-incorporation efficace de l'ETH et du booster a dû surmonter de nombreuses difficultés liées à des problèmes de solubilité, de cristallisation et de biodisponibilité. Nos NPs en PLA et en pCD ont montré leur capacité de co-encapsuler efficacement les deux molécules et tout particulièrement celles en pCD. Elles incorporent les médicaments à la fois dans les cavités des CD et dans des microdomaines hydrophobes. Les NPs en pCD, non toxiques après administration pulmonaire répétée de fortes doses, ont été administrés in vivo par voie endotrachéale directement au site d'infection. Elles ont permis une diminution de 3-log de la charge bactérienne pulmonaire des animaux infectés après seulement 6 administrations. De même, l'incorporation de PIP a été confrontée à des défis liés à la cristallisation de PIP et à sa libération incontrôlée. Malheureusement, le PIP présentait une faible affinité pour tous les matériaux polymériques étudiés et son encapsulation physique était infructueuse. Ainsi, une approche alternative a été développée en couplant le PIP au PCL via une réaction sans catalyseur initiée par le médicament. Le conjugué PCL-PIP se auto-assemble en forme de NPs avec une charge en PIP de 27%. Cependant, le PCL-PIP n'a pas pu être dégradé in vitro, mais l’approche de synthèse de conjugués est séduisante pour obtenir de particules stables et avec un contenu important en PIP.La compréhension approfondie de la structure et de la composition du noyau et de la couronne des nanostructures contenant une ou deux molécules actives est cruciale pour leur optimisation. Les deux derniers chapitres sont donc consacrés à l'application innovante de l'AFM-IR, une méthode nanospectroscopique originale combinant la microscopie à force atomique (AFM) avec la spectroscopie infrarouge (IR), à l'analyse chimique des NPs en PLGA ou à leur détection sans marquage après internalisation dans les cellules.L’AFM-IR est capable de fournir une caractérisation chimique à l'échelle nanométrique (résolution ~10 nm). Une avancée majeure du travail est l'application du mode tapping permettant l'investigation individuelle de chaque NP. Le signal IR spécifique des composants des NPs a été utilisé pour appréhender la composition chimique de leur cœur et couronne ainsi que pour localiser précisément le médicament. De plus, l'AFM-IR en mode contact a permis pour la première fois la localisation sans marquage et l'identification chimique des NP à l'intérieur des cellules. Ce travail ouvre la voie à d'innombrables applications de cette technique dans le domaine de la nanomedecine
The treatment of intracellular infections is very challenging given the ability of bacteria to “hide” inside the cells of the host, especially the ones of the immune system, thus hampering the action of many antimicrobial agents. The battle against these bacteria has been further exacerbated by the increasing diffusion of antimicrobial resistant strains. In this frame, nanoparticles (NPs) are a very promising strategy to overcome the limitations of free antimicrobial agents by administering them in an optimized manner.This PhD work, performed as part of the European Project ITN Cyclon Hit, aimed at the development and advanced characterisation of antibiotic-loaded biodegradable and biocompatible NPs made of poly (lactic acid) (PLA), poly (lactic-co-glycolic) (PLGA) and polycaprolactone (PCL) or of polymerised cyclodextrins (pCDs).The first two chapters are dedicated to the encapsulation of powerful but challenging drugs in polymeric NPs. Firstly, these carriers were employed for the simultaneous delivery of a potent drug combination recently discovered, ethionamide (ETH) and its booster, for tuberculosis therapy. Secondly, they were used to address the challenges related to the incorporation of a first-generation quinolone, pipemidic acid (PIP), with the aim of optimising its intracellular delivery in infections such as salmonellosis.The efficient co-incorporation of ETH and booster had to overcome several technological barriers. These drugs presented solubility, crystallisation and bioavailability-related problems which were overcome thanks to the developed NPs. Our engineered PLA and pCD NPs were both able to efficiently co-encapsulate the two molecules. Among the in depth-characterised formulations, pCDs NPs displayed the best physico-chemical properties and were shown to host the drugs both in the CD cavities and in confined spaces inside NPs crosslinked polymer. The pCD NPs were administered in vivo by endotracheal route directly to the infection site. Empty NPs were shown non-toxic after repeated pulmonary administration of high doses. Moreover, loaded pCD NPs led to a 3-log decrease in the pulmonary bacterial load of infected animals after only 6 administrations. Similarly, the incorporation of PIP faced challenges mainly related to PIP crystallization and burst release. Unfortunately, PIP displayed poor affinity for all the studied polymeric materials and its physical encapsulation was unsuccessful. Thus, an alternative approach was developed by coupling PIP to PCL by using an original catalyst-free drug-initiated reaction. The PCL-PIP conjugate self-assembled in NPs with up to 27 wt% PIP which were thoroughly characterised. However, the conjugate couldn’t be enzymatically degraded. With the design of novel PCL-PIP conjugates, this self-assembly approach could represent a promising strategy.The deep understanding of the structure and composition of complex core-corona nanocarriers containing one or two active molecules is crucial for their optimisation. The last two chapters are devoted to the innovative application of AFM-IR, an original nanospectroscopic method combining atomic force microscopy (AFM) with infrared (IR) spectroscopy, to the chemical analysis of PLGA NPs or to their label-free detection after cell internalisation.AFM-IR is able to provide chemical characterisation at the nanometer scale (resolution ~10nm). One main breakthrough here is the application of the recently developed tapping mode allowing the investigation of single polymeric NPs. The specific IR signal of NPs constituents was used to unravel the chemical composition of their core and corona as well as to precisely locate the drug. Moreover, the AFM-IR in contact mode enabled for the first time the label-free localisation and unambiguous chemical identification of NPs inside cells using the polymer IR specific response as a fingerprint. This work paves the way for countless application of this technique in the field of drug delivery

[EN] In the present Doctoral Thesis, different strategies to improve functional properties of starch films for food packaging applications were analysed: study of the effect of amylose:amylopectin ratio, blend with other polymers poly(vinyl alcohol) (PVA), and incorporation of different fillers (rice bran and cellulose nanocrystals-CNCs) and antimicrobial agents (neem oil-N, oregano essential oil-O and silver nanoparticles-AgNPs). Likewise, a biodegradation study of the films as affected by antimicrobials was carried out. Among different starches with distinct amylose:amylopectin ratio, pea starch was selected with higher values of this ratio. The high content of amylose gave rise to stiffer and more resistant to fracture films, with lower oxygen permeability, which decreased during storage. Pea starch-PVA (S-PVA) blend films represented a good strategy to improve film properties without a notable increase in cost. The PVA addition led to films which were less water soluble and not as sensitive to water sorption, more extensible and resistant than neat starch films, while they maintained the high oxygen barrier and provided stability to the matrix during ageing. In this sense, S-PVA ratios near to 1:1 are recommendable. When rice bran with the smallest particle size was used as filler, it improved the elastic modulus of the films, but reduced the film stretchability and barrier properties, due to the enhancement of the water binding capacity and the introduction of discontinuities in the matrix. Incorporation of CNCs into S-PVA films, as a reinforcing material, enhanced phase separation of polymers, did not imply changes in water vapour barrier of the films but improved the film mechanical performance: they became stiffer and more stretchable, while crystallization of PVA was partially inhibited. Silver loaded S-PVA films exhibited antimicrobial activity against the fungi (Aspergillus niger and Penicillium expansum) and bacteria (Listeria innocua and Escherichia coli) genera, depending on the silver concentration. Silver nanoparticles provoked notable changes in the film colour and transparency but no relevant changes in the other physical properties. Silver was completely delivered to aqueous simulants within the firsts 60 minutes of contact, but the film release capacity drastically decreased in the non-polar simulant, where the overall migration limit for food contact packaging materials (60 mg/Kg simulant) was accomplished. So, the use of the developed films as food packaging materials should be restricted to fat-rich foodstuffs. The incorporation of oregano essential oil (O), as antimicrobial agents, into the S-PVA matrix gave rise to active films against bacteria and fungi, whereas films containing neem oil (N) were not effective. Higher O concentration in the films was required to observe antifungal effect with respect to the antibacterial activity. Oils did not notably affect water sensitivity and water barrier properties of the films, but at high ratio, oils implied weaker film networks, affecting their mechanical performance. Blend of starch with PVA significantly improved PVA biodegradation and disintegration behaviour. Incorporation of silver species to S-PVA films slowed down their biodegradation kinetics while increased their degradation ratio. However, no significant effect of O was observed on the biodegradability indices despite the antimicrobial activity detected for this oil. N even promoted biodegradation of S-PVA films. Finally, a food application of biopolymer coatings was studied, using chitosan and essential oils (oregano or rosemary) to prevent weight loss and fungal development of semi-hard goat's milk cheeses. Coating were highly effective at reducing fungal growth and cheese weight loss while slightly reduced lipolytic and proteolytic activities in the cheese. Sensory evaluation revealed that the cheeses double coated with chitosan-oregano oil were the best evaluated in terms of aroma and flavour.
[ES] En la Tesis Doctoral, se han analizado diferentes estrategias para mejorar las propiedades funcionales de películas de almidón para aplicaciones de envasado de alimentos: el estudio del ratio amilosa:amilopectina, mezclas con otros polímeros (alcohol de polivinilo-PVA), y la incorporación de diferentes refuerzos (salvado de arroz y nanocristales de celulose-CNCs) y agentes antimicrobianos (aceite de neem-N, aceite esencial de orégano-O y nanopartículas de plata-AgNPs). También se realizó un estudio de biodegradación de las películas observando el efecto de los antimicrobianos. Entre los diferentes almidones con distinto ratio amilosa:amilopectina, se seleccionó el almidón de guisante con altos valores de amilosa. El alto contenido en amilosa dio lugar a películas más rígidas y resistentes a la fractura, con baja permeabilidad al oxígeno, la cual disminuyó durante el almacenamiento. Las películas de mezcla de almidón-PVA (S-PVA) representaron una buena estrategia para mejorar las propiedades de las películas sin incrementar notablemente el precio. La adición de PVA dio lugar a películas menos solubles en agua, menos sensibles a su absorción y más extensible y resistentes que las de almidón puro, mientras mantuvieron alta barrera al oxígeno y dieron estabilidad a la matriz durante el envejecimiento. Son recomendables los ratios de S-PVA cercanos a 1:1. El salvado de arroz con menor tamaño de partícula, mejoró el modulo elástico de las películas, pero su extensibilidad y propiedades barrera empeoraron debido a la mejor capacidad de retención de agua y a las discontinuidades introducidas. La incorporación de CNCs en las películas de S-PVA incrementó la separación de fases de los polímeros, sin cambios en la permeabilidad al vapor de agua, y mejoró las prestaciones mecánicas: películas más rígidas y extensibles, mientras que inhibió parcialmente la cristalización del PVA. Las películas de S-PVA con partículas de plata exhibieron actividad antimicrobiana contra dos hongos y dos bacterias, dependiendo de la concentración de plata. Las AgNPs provocaron cambios en el color de las películas así como en su transparencia. La plata fue completamente liberada en los primeros 60 minutos en contacto con simulantes acuosos, sin embargo la liberación disminuyó en simulantes no polares, donde se cumple el límite de migración global. Por lo tanto, el uso de las películas desarrolladas para envasado de alimentos debe ser restringido a productos alimenticios ricos en grasas. La incorporación de O en la matriz de S-PVA dio lugar a películas activas contra bacterias y hongos, mientras que las películas con N no fueron efectivas. Se necesitaron concentraciones más altas de O en las películas para observar un efecto antifúngico respecto a la actividad bactericida. Los aceites no afectaron notablemente la sensibilidad ni las propiedades barreras al agua de las películas, aunque la mayor proporción de aceite dio lugar a películas con una red más débil, afectando sus prestaciones mecánicas. La mezcla de S con PVA mejoró significativamente el comportamiento de biodegradación y desintegración de las películas. La incorporación de AgNPs a películas de S-PVA disminuyó su cinética de biodegradación mientras aumentó su ratio de degradación. La adición de O no presentó efecto significativo en el índice de biodegradación a pesar de la actividad antimicrobiana detectada. El N incluso mejoró la biodegradación de las películas. Finalmente, se realizó una aplicación de recubrimientos basados en biopolímeros, usando quitosano-CH y aceite esencial de orégano o romero para evitar la pérdida de peso y el desarrollo de hongos en quesos de cabra semicurados. Los recubrimientos fueron efectivos en la reducción del crecimiento fúngico y la pérdida de peso de los quesos, mientras que la actividad lipolítica y proteolítica ligeramente disminuyó. El análisis sensorial reveló q
[CAT] S'han analitzat diferents estratègies per a millorar les propietats funcionals de pel·lícules de midó per a aplicacions d'envasat d'aliments: l'estudi de la ràtio amilosa:amilopectina, mescles amb altres polímers (alcohol polivinílic-PVA), i la incorporació de diferents reforços (segó d'arròs i nanocristalls de celulosa-CNCs) i agents antimicrobians (oli de neem-N, oli essencial d'orenga-O i nanopartícules de plata). També es va fer un estudi de biodegradació de les pel·lícules per observar l'efecte dels antimicrobians. Entre els diferents midons amb distint ràtio amilosa:amilopectina, es va seleccionar el midó de pèsol amb alts valors d'amilosa. L'alt contingut en amilosa va donar lloc a pel·lícules més rígides i resistents a la fractura, amb baixa permeabilitat a l'oxigen, la qual va disminuir durant l'emmagatzemament. Les pel·lícules de mescla de S-PVA van representar una bona estratègia per a millorar les propietats de les pel·lícules sense incrementar notablement el preu. L'addició de PVA va donar lloc a pel·lícules menys solubles en aigua, menys sensibles a la seua absorció i més extensible i resistents que les de midó pur. A més van mantenir l'alta barrera a l'oxigen i van donar estabilitat a la matriu durant l'envelliment. Son recomanables les ràtios de S- PVA pròxims a 1:1. El segó d'arròs amb menor tamany de partícula, va millorar el mòdul elàstic de les pel·lícules, però la seua extensibilitat i propietats barrera van empitjorar. La incorporació de CNCs en les pel·lícules de S-PVA, va incrementar la separació de fases dels polímers, sense implicar canvis en la permeabilitat al vapor d'aigua, però millorant les prestacions mecàniques: pel·lícules més rígides i extensibles, al mateix temps que es va inhibir parcialment la cristal·lització del PVA. Les pel·lícules de S-PVA amb partícules de plata van exhibir activitat antimicrobiana front a dos de fongs i dos bacteris, depenent de la concentració de plata. Les AgNPs van provocar canvis en el color de les pel·lícules així com en la seua transparència. La plata va ser completament alliberada en els primers 60 minuts en contacte amb simulants aquosos. La capacitat d'alliberament va disminuir en simulants no polars, on es complix el límit de migració global. Per tant, l'ús de les pel·lícules desenvolupades per a l'envasat d'aliments ha de ser restringit a productes alimentaris rics en greixos. La incorporació d'O en la matriu de S-PVA va donar lloc a pel·lícules actives front a bacteris i fongs. Pel contrari les pel·lícules amb N no van ser efectives. Van ser necessàries concentracions més elevades d'O en les pel·lícules per a observar un efecte antifúngic respecte a l'activitat bactericida. Els olis no van afectar la sensibilitat a l'aigua ni les propietats barrera a l'aigua de les pel·lícules, però la major proporció d'oli va donar lloc a pel·lícules amb una xarxa més dèbil, afectant les seues prestacions mecàniques. La mescla de S amb PVA va millorar significativament el comportament de biodegradació i desintegració. La incorporació de partícules de plata a pel·lícules de S-PVA va disminuir la seua cinètica de biodegradació mentre que va augmentar la seua ràtio de degradació. No obstant això, l'addició d'O no va presentar un efecte significatiu en els índexs de biodegradació a pesar de l'activitat antimicrobiana detectada. L'N fins i tot va millorar la biodegradació de les pel·lícules de S-PVA. Finalment, es va realitzar una aplicació de recobriments basats en biopolímers, fent ús de quistosà-CH i olis essencials de orenga i romer per evitar la pèrdua de pes i el desenvolupament de fongs en formatges de cabra semi-curats. Els recobriments van ser altament efectius en la reducció del creixement fúngic i la pèrdua de pes dels formatges. L'activitat lipolítica i proteolítica en els formatges va disminuir suaument. L'anàlisi sensorial va revelar que els form
Cano Embuena, AI. (2015). Different strategies to improve the functionality of biodegradable films based on starch and other polymers [Tesis doctoral no publicada]. Universitat Politècnica de València. https://doi.org/10.4995/Thesis/10251/55383
TESIS
Premiado

World population is continuously growing, as well as the influence we have on the ecosystem’s natural equilibrium. Moreover, such growth is not homogeneous and it results in an overall increase of older people. Humanity’s activity, growth and aging leads to many challenging issues to address: among them, there are the spread of suddenly and/or chronic diseases, malnutrition, resource pressure and environmental pollution. Research in the novel field of biodegradable soft robotics and electronics can help dealing with these issues. In fact, to face the aging of the population, it is necessary an improvement in rehabilitation technologies, physiological and continuous monitoring, as well as personalized care and therapy. Also in the agricultural sector, an accurate and efficient direct measure of the plants health conditions would be of help especially in the less-developed countries. But since living beings, such as humans and plants, are constituted by soft tissues that continuously change their size and shapes, today’s traditional technologies, based on rigid materials, may not be able to provide an efficient interaction necessary to satisfy these needs: the mechanical mismatch is too prohibitive. Instead, soft robotic systems and devices can be designed to combine active functionalities with soft mechanical properties that can allow them to efficiently and safely interact with soft living tissues. Soft implantable biomedical devices, smart rehabilitation devices and compliant sensors for plants are all applications that can be achieved with soft technologies. The development of sophisticated autonomous soft systems needs the integration on a unique soft body or platform of many functionalities (such as mechanical actuation, energy harvesting, storage and delivery, sensing capabilities). A great research interest is recently arising on this topic, but yet not so many groups are focusing their efforts in the use of natural-derived and biodegradable raw materials. In fact, resource pressure and environmental pollution are becoming more and more critical problems. It should be completely avoided the use of in exhaustion, pollutant, toxic and non-degradable resources, such as lithium, petroleum derivatives, halogenated compounds and organic solvents. So-obtained biodegradable soft systems and devices could then be manufactured in high number and deployed in the environment to fulfil their duties without the need to recover them, since they can safely degrade in the environment. The aim of the current Ph.D. project is the use of natural-derived and biodegradable polymers and substances as building blocks for the development of smart composite materials that could operate as functional elements in a soft robotic system or device. Soft mechanical properties and electronic/ionic conductive properties are here combined together within smart nanocomposite materials. The use of supersonic cluster beam deposition (SCBD) technique enabled the fabrication of cluster-assembled Au electrodes that can partially penetrate into the surface of soft materials, providing an efficient solution to the challenge of coupling conductive metallic layers and soft deformable polymeric substrates. In this work, cellulose derivatives and poly(3-hydroxybutyrate) bioplastic are used as building blocks for the development of both underwater and in-air soft electromechanical actuators that are characterized and tested. A cellulosic matrix is blended with natural-derived ionic liquids to design and manufacture completely biodegradable supercapacitors, extremely interesting energy storage devices. Lastly, ultrathin Au electrodes are here deposited on biodegradable cellulose acetate sheets, in order to develop transparent flexible electronics as well as bidirectional resistive-type strain sensors. The results obtained in this work can be regarded as a preliminary study towards the realization of full natural-derived and biodegradable soft robotic and electronic systems and devices.

The goal of this project was to develop a non-toxic amphiphilic diblock copolymer nanoparticulate drug delivery system that will solubilize paclitaxel (PTX) and retain the drug in plasma. Methoxy poly(ethylene glycol)-block-poly(ε-caprolactone) (MePEG-b-PCL) diblock copolymers loaded with PTX were characterized and their physicochemical properties were correlated with their performance as nanoparticulate drug delivery systems. A series of MePEG-b-PCL was synthesized with PCL blocks ranging from 2-104 repeat units and MePEG blocks of 17, 44 or 114 repeat units. All copolymers were water soluble and formed micelles except MePEG₁₁₄-b-PCL₁₀₄, which was water insoluble and formed nanospheres. Investigation of the effects of block length on the physicochemical properties of the nanoparticles was used to select appropriate copolymers for development as PTX nanoparticles. The critical micelle concentration, pyrene partition coefficient and diameter of nanoparticles were found to be dependent on the PCL block length. Copolymers based on a MePEG molecular weight of 750 g/mol were found to have temperature dependent phase behavior. Relationships between the concentration of micellized drug and the compatibility between the drug and core-forming block, as determined by the Flory-Huggins interaction parameter, and PCL block length were developed. Increases in the compatibility between PCL and the drug, as well as longer PCL block lengths resulted in increased drug solubilization. The physicochemical properties and drug delivery performance characteristics of MePEG₁₁₄-b-PCL₁₉ micelles and MePEG₁₁₄-b-PCL₁₀₄ nanospheres were compared. Nanospheres were larger, had a more viscous core, solubilized more PTX and released it slower, compared to micelles. No difference was seen in the hemocompatibility of the nanoparticles as assessed by plasma coagulation time and erythrocyte hemolysis. Micellar PTX had an in vitro plasma distribution similar to free drug. The majority of micellar PTX associated with the lipoprotein deficient plasma fraction (LPDP). In contrast, nanospheres were capable of retaining more of the encapsulated drug with significantly less PTX partitioning into the LPDP fraction. In conclusion, although both micelles and nanospheres were capable of solubilizing PTX and were hemocompatible, PTX nanospheres may offer the advantage of prolonged blood circulation, based on the in vitro plasma distribution data, which showed that nanospheres retained PTX more effectively.

Chemical catalysts are divided into two traditional categories: homogeneous and heterogeneous catalysts. Although homogeneous (molecular) catalysts tend to have high activity and selectivity, their wide application is hampered by the difficulties in catalyst separation. In contrast, the vast majority of industrial scale catalysts are heterogeneous catalysts based on solid materials. Immobilized catalysts, combining the advantages of homogeneous and heterogeneous catalysts, have developed into an important field in catalysis research. This dissertation presents synthesis, characterization and evaluation of several novel immobilized catalysts. In the first part, MNP supported aluminum isoproxide was developed for ROP of Є-caprolactone to achieve facile magnetic separation of catalysts from polymerization system and reduce toxic metal residues in the poly(caprolactone) product. Chapter 3 presents a silica coated MNP supported DMAP catalyst that was synthesized and displayed good activity and regio-selectivity in epoxide ring opening reactions. In Chapter 4, hybrid sulfonic acid catalysts based on polymer brush materials have been developed. The unique polymer brush architecture permits high catalyst loadings as well as easy accessibility of the active sites to be achieved in this catalytic system. In Chapter 5, aminopolymer-silica composite supported Pd catalysts with good activity and selectivity were developed for the selective hydrogenation of alkynes. In this case, the aminopolymer composite works as a stabilizer for palladium nanoparticles, as well as a modifier to tune the catalyst selectivity. All in all, the general theme of the thesis is developing new immobilized catalysts with improved activity/selectivity as well as easy separation via rational catalyst design.

$EVWUDFW Application of radionuclides due to their unique nature is still attracting the interest of professionals from different branches of science. In this thesis, the unifying topic is the utilization of various radioactive labels on polymeric carriers for biomedicinal purposes. In the introductory part, the area of use of the investigated substances - in terms of their possible application in nuclear medicine, is presented to the reader. The experimental work is divided into three main sections dealing with synthesis and studying properties of appropriate selected polymers, their metal chelates and radiolabeling. First part of the study describes development of new strategies for labeling of polymeric carriers with radiometal cations and radioiodine, whereas in the following sections of the study synthesis and investigation of biocompatible polymer materials is discussed. New possible approaches in the diagnostics and therapy of cancer by using polymeric chelates as carriers are presented as well as the use of macroporous metal chelating polymer beads as potential therapeutics intended for radioembolization of liver malignancies and for the therapy of Wilson's disease. Investigated polymers have been shown to be non-toxic, to have good in vitro stability and to have the ability to be radiolabeled in...

碩士
國立臺灣大學
化學研究所
101
Amphiphilic block polycarbonates have received significant attention in designing a variety of therapeutics and diagnosis owing to their biocompatibility, and tunable mechanical properties. Herein we demonstrate a biodegradable amphiphilic polycarbonate (MTC-MPA-3HF) bearing ROS-reactive pendent groups as a new biomaterial to sense ROS in physiological environment. Before the treatment of stimuli, MTC-MPA-3HF assembles into a stable nanoparticle in aqueous solution resulting ESIPT-dominated green fluorescence in hydrophobic milieu, whereas MTC-MPA-3HF may disassemble upon the treatment of ozone- or hydrogen peroxide emitting ESICT-dominated blue fluorescence (Figure A). Correspondingly, two polymeric structures decorating with different reactive triggers in more hydrophobic block to sense ozone and hydrogen peroxide were designed and synthesized. O3-responsive polymer (MTC-MPA-3HF-O3) employs MTC-Obutene to serve as the ozone-reactive moiety as well as to confer the required hydrophobicity, while H2O2-responsive polymer (MTC-MPA-3HF-H2O2) having MTC-O3dCHBE or MTC-OArBE to serve as the H2O2-reactive group (Figure B). Both polymers feature the same hydrophilic unit, MTC-OmDEG, and the environmentally-sensitive bis-MPA-3HF fluorophore as an initiator. The fluorescence behaviors and the morphology of the particles were studied by fluorescence spectroscopy and transmission electron microscope, respectively, before and after the treatment of stimuli. The results showed that in the case of MTC-MPA-3HF-O3, ozone seems to react with bis-MPA-3HF much faster than MTC-Obutene resulting in fluorescence quenching without manifesting size change. However, MTC-MPA-3HF-O3 displayed an excellent specificity to the lipase from Candida Antarctica. Either the skeleton or the pendent of the polymer was presumably hydrolyzed to give the fluorescence color change, rendering the application of MTC-MPA-3HF-O3 in antibacterial therapy quite feasible. In the case of MTC-MPA-3HF-H2O2 with alkyl boronic ester, nanoparticles became 500-fold larger with ratiometric fluorescence readout upon reacting with hydrogen peroxide. It is worthwhile to note that replacing alkyl boronic ester with aryl boronic ester gave even better ratiometric fluorescence signals. In conclusion, we have successfully developed a series of biodegradable amphiphilic polycarbonates bearing ozone- or hydrogen peroxide-reactive groups that may disassemble upon exposure to each targeted stimuli with concomitant fluorescence color change. Furthermore, taking advantages of ester-rich MTC-MPA-3HF, it is feasible to detect the bacterial lipase by monitoring the ratiometric fluorescent readout. These materials can be potentially used in delivering therapeutics or serve as diagnostic agents.

A magnetic target drug delivery system consisting of biodegradable polymeric microspheres (poly D, L-lactic acid) loaded with magnetite nanoparticles (10-100 nm) and anticancer drug (paclitaxel) was studied. The magnetite nanoparticles were synthesized by chemical precipitation. The as-synthesized magnetite nanoparticles were subsequently introduced into a mixture of polymer magnetic polymeric composite particles were investigated and further correlated with the reaction parameters. It was found that the size and characteristics of the polymeric composite particles depended on the viscosity of the polymer solution. Preliminary drug release experiments showed that the loaded drug was released with the degradation of the polymer. The release rates could be enhanced by an oscillating external magnetic field.
Singapore-MIT Alliance (SMA)

no
This review summarises the recent developments in the synthesis and applications of polymers derived from malic acid. There has been an increased interest in the design of sustainable and biodegradable polymers as a result of the drive to use renewable feedstocks as an alternative to petrochemicals in addition to their significant potential in biomedical applications. Synthetic strategies to access polymers from malic acid based on both condensation and ring-opening polymerization, across a broad range of conditions, are reviewed along with their advantages and limits. The role that such materials are studied for in biomedical applications is discussed, and their environmental impact based on the biodegradability of the malic polymer backbone is outlined.
The Royal Society, EPSRC, BBSRC

Charles University Faculty of Pharmacy in Hradci Králové Department of Pharmaceutical Technology Consultant: PharmDr. Ondřej Holas, PhD. Student: Petra Staňková Title of Thesis: Pharmaceutical applications of polyesters as drug nanocarriers Nanoparticles are nowadays intensively studied and perspective type of a drug carrier. Its potential is based on a possibility of targeted drug delivery and controlled drug release. The theoretical part is about nanoparticles types, polymers derived from α-hydroxyacids (PLA, PGA, PLGA). Focus is given on methods of nanoparticles preparation: dispersion of preformed polymers or the polymerization of monomers. The modification of particles surface and practical use of nanomedicine in healthcare are described in other chapters. The research in experimental part is focused on the influence of different types of PLGA and their weighing on the size, polydispersity, nanoparticles zeta potential and encapsulation efficiency of rhodamine B. Nanoparticles were prepared by nanoprecipitation method or by solvent evaporation method. The Zetasiser ZS 90 device was used to measure the size of the nanoparticles and to measure zeta potential. The result of the research shows the most suitable weighing for creation of nanoparticles is 25 mg. The samples of this weighing show a...

Charles University in Prague, Faculty of Pharmacy in Hradec Králové Department of: Pharmaceutical technology Consultant: PharmDr. Ondřej Holas, Ph.D. Student: Silvia Bacskaiová Title of Thesis: Biodegradable polymeric nanoparticles preparation The present scientific progress in an important rate conduces to nanomedicine development, which aims to reengineering of cancer pharmacotherapy and other substantial diseases. The main intention of this graduation thesis is the study of surface-active chemical's effects on the final properties of nanoparticles. The theoretical part is focused on the nanoparticles likedrugs vehicles, synthesis of biodegradable nanoparticles, themselves applications in the cancer therapy, diseases accompanied by inflammations, vaccination and for another different purposes. The theoretical part also contents characterization of active and passive goal- directed distribution by diseases, microencapsulations, synthesis of nanoparticles by polymerization and from previously synthesized polymers and definition of physical- chemical properties of nanoparticles. The experimental part is more extensive and concerned with the optimalization of reaction conditions of nanoparticles synthesis, with selection of advisable concentracion and type of surfactant. In the experimental part were...

Charles University, Faculty of Pharmacy in Hradec Králové Department of: Pharmaceutical Technology Consultant: PharmDr. Ondřej Holas, Ph.D. Student: Michal Bárta Title of Thesis: Preparation of pharmaceutical nanoparticles: optimalization process Pharmaceutical, polymer nanoparticles besides others work as well as a drug carriers. Their uniqueness is not only because of their subcelular size, but as well because of the biodegradability and biocompatibility they hold. The benefit of these nanoparticiples is the possibility of creation of selective naoparticiples which are able to controle long-term release. The formulation of polymer nanoparticles can be reached within methods using preformed polymer or with polymerization of monomers. The main goal of my thesis was to optimize the production process of polymer nanoparticles and the observation of the solvent influences. The methods used for this reaserch were evaporation method and nanoprecipitation. Granulometric and electrical characteristics of particles were measured with Zetasizer ZS 90. Measurements have prooved, that it is preferable to use the nanoparticle method for the prepartion of the the small nanoparticles with low polydispersity and sufficient stability. From the results of the granulometric analysis of nanoparticles made by the...

The thesis outlines possible medical applications of soft matter assemblies as nanotechnology based systems as well as their potential in the emerging field of nanomedicine. Nanomedicine can be defined as the investigation area encompassing the design of diagnostics and therapeutics at the nanoscale, including nanobots, nanobiosensors, nanoparticles and other nanodevices, for the remediation, prevention and diagnosis of a variety of illnesses. The ultimate goal of nanomedicine is to improve patient quality-of-life. Because nanomedicine includes the rational design of an enormous number of nanotechnology-based products focused on miscellaneous diseases, a variety of nanomaterials can be employed. Therefore, the thesis is driven by a focus on recent advances in the manufacture of soft matter-based nanomedicines specifically designed to improve cancer diagnostics and chemotherapy efficacy. It will in particular highlight liposomes, polymer-drug conjugates, drug- loaded block copolymer micelles and biodegradable polymeric nanoparticles, emphasizing the current investigations and potential novel approaches towards overcoming the remaining challenges in the field as well as a brief overview of formulations that are in clinical trials and marketed products. Based on vehicle-related and...