Vaccines have become an important approach for the treatment of cancer. Cancer vaccines help the immune system to detect and eradicate tumor cells. Also, cancer vaccines are designed to stimulate an effective immune response that can create long-term immune memory to prevent tumor recurrence. This treatment approach involves the administration of a vaccine comprising or encoding an antigen and can often be combined with an adjuvant to further promote the immune response. The goal of this research was to study the effect of the polyanhydride composition of prophylactic cancer vaccine formulations on the tumor-specific immune response. To achieve this goal, three different amphiphilic polyanhydride copolymers were generated comprising different ratios of 1,6-bis-(p-carboxyphenoxy)-hexane (CPH) and 1,8-bis-(p-carboxyphenoxy)-3,6-dioxaoctane (CPTEG) or sebacic anhydride (SA) monomers. These copolymers were used to fabricate particles encapsulating a model antigen, ovalbumin (OVA), using a double emulsion solvent evaporation technique. The ability of the three different compositions of amphiphilic polyanhydride copolymers (50:50 CPTEG:CPH, 20:80 CPTEG:CPH, and 20:80 CPH:SA) encapsulating OVA to elicit immune responses was investigated. Further, the impact of soluble unmethylated oligodeoxynucleotides containing deoxycytidyl-deoxyguanosine dinucleotides (CpG ODN), an immunologic adjuvant, on the immune response to the three formulations was also studied. The immune response to cancer vaccines was measured after treatment of C57BL/6J mice with two subcutaneous injections, seven days apart, of 50 μg OVA encapsulated in particles composed of different polyanhydride copolymers with or without 25 μg CpG ODN. In vivo studies showed that 20:80 CPTEG:CPH particles encapsulating OVA significantly stimulated the highest level of CD8+ T lymphocytes, generated the highest serum titers of OVA-specific IgG antibodies, and produced longer survival in comparison to formulations involving the other polyanhydride copolymers. The results also revealed that supplementing the vaccine formulations with CpG ODN did not enhance the immunogenicity of OVA. These results accentuate the crucial role of the copolymer composition of polyanhydrides in stimulating the immune response and improving cancer vaccine efficacy.

Mémoire numérisé par la Direction des bibliothèques de l'Université de Montréal.

Hidrogéis compreendem uma importante classe de materiais poliméricos adequados à aplicação como curativos de feridas e queimaduras. A estrutura tridimensional hidrofílica dos hidrogéis permite que estes mantenham a umidade ideal no leito das feridas, absorvam o exsudato e não causem danos ao novo tecido durante as trocas dos curativos. No caso dos hidrogéis, essas trocas podem ser menos frequentes. Além disso, curativos que auxiliem na remoção de tecidos necrosados e ainda sejam capazes de oferecer tratamentos extras que acelerem o processo de cicatrização são desejáveis. Este trabalho apresenta a produção de materiais à base de hidrogel capazes de auxiliar neste processo de diferentes maneiras. Primeiramente, são apresentados hidrogéis formados a partir de nanofibras de poli(N-vinil-2-pirrolidona) (PVP) produzidas por eletrofiação, seguido da reticulação através da utilização de radiação UV-C ou reação de Fenton. A utilização da eletrofiação como técnica auxiliar na formação dos hidrogéis permitiu o controle da porosidade através da formação de fibras de diferentes diâmetros. A evidência de tal propriedade foi constatada através da produção de materiais que apresentam diferentes perfis de liberação da proteína modelo albumina de soro bovino (BSA). O hidrogel de PVP nanoestruturado foi capaz de liberar e manter a atividade da colagenase, uma importante enzima aplicada no tratamento de feridas via desbridamento enzimático, durante as 48 horas em que foi avaliado. Além disso, hidrogéis bactericidas nanoestruturados foram produzidos a partir de nanocompósitos de PVP e nanopartículas de prata (AgNP) produzidos por eletrofiação. Esses hidrogéis apresentaram propriedades térmicas semelhantes aos hidrogéis sem AgNP, diminuindo, contudo, a sua capacidade de intumescimento. Esses hidrogéis mostraram-se ativos contra bactérias gram-positivas e gram-negativas a partir de 100 ppm de AgNPs. Adicionalmente, foi estudada a formação de um hidrogel modelo composto PVP/AgNP/Imidazol, almejando-se a produção de um material bactericida-fungicida a base de hidrogel. Este hidrogel apresentou atividade conta três espécies de Candida a partir de 500 ppm de imidazol no material. Embora exista a formação de um complexo estável entre AgNP e Imidazol, cálculos teóricos e a constatação da atividade fungicida corroboram com o fato de que derivados imidazólicos podem ser liberados a partir deste hidrogel híbrido. A produção de hidrogéis físicos compostos por blendas de PVP/Polianidridos sintetizados a partir de derivados de hidroxicinamatos e ácido salicílico, capazes de liberar moléculas de interesse biológico quando parcialmente degradados hidroliticamente, também é descrita neste trabalho. Os resultados indicam que interações hidrofóbicas entre a PVP e os polianidridos sintetizados podem ser responsáveis pela formação dos hidrogéis físicos e pela miscibilidade das blendas produzidas. Os hidrogéis físicos de PVP/Polianidridos foram obtidos na forma de filmes por evaporação do solvente. Micro- e nanofibras também foram obtidas por eletrofiação. Desta maneira, o presente trabalho contribui com o desenvolvimento de uma geração de curativos multifuncionais aplicados no tratamento de feridas crônicas e queimaduras. / Hydrogels comprise an important class of polymeric materials that finds application as wound and burn dressings. The hydrophilic three-dimensional structure of hydrogels helps to provide the ideal humidity at the wound bed, to remove exsudates and to prevent damages to the new tissue during dressing substitution. Furthermore, these wound dressings are able to remove necrotic tissues and, therefore, capable to offer extra treatments that would benefit the healing processes. This work describes the production of hydrogel based materials that are able to act in wound healing by different ways. First, it is presented hydrogels composed of poly(N-vinyl-2-pyrrolidone) (PVP) nanofibers produced by electrospinning, followed by its crosslinking using UV-C radiation or Fenton reaction. The use of electrospinning in the hydrogel formation allowed porosity control by obtaining fibers of different diameters. This was evidenced by achieving materials that present different release profiles of the model protein bovine serum albumin (BSA). The nanostructured PVP hydrogel was capable of releasing and maintaining collagenase activity during 48 hour of evaluation. This is an important enzyme that find application in wound healing based on enzymatic debridement. Moreover, nanostructured bactericidal hydrogels were produced from PVP and silver nanoparticles (AgNP) composite through electrospinning, resulting in hydrogels with thermal properties similar to those hydrogels without AgNP, decreasing its swelling ability. These hydrogels were active against gram-positives and gram-negatives bacteria starting from 100 ppm of AgNP. In addition, the production of a model hydrogel composed by PVP/AgNP/Imidazole was investigated, aiming at a bactericidal-fungicidal hydrogel based material. This hydrogel was active against three Candida having 500 ppm of imidazole into the structure. In spite of the formation of a stable complex between AgNP and imidazole, theoretic calculations and the observed fungicidal activity corroborate with the fact that imidazoles derivatives can be released from this hybrid hydrogel. Physical hydrogels composed of PVP/Polyanhydrides blends were synthesized from hydroxycinammates derivatives and salicylic acid. These materials which were capable of releasing molecules with biological potential upon hydrolysis, are also described in this work. The results indicate that hydrophobic interactions between PVP and the synthesized polyanhydrides could be responsible for the hydrogel formation and blend miscibility as well. PVP/Polyanhydride physical hydrogels were obtained from cast films. Micro- and nanofibers were also obtained by electrospinning. Thus, the present work contributes with the development of the new generation of smart dressings for wound and burn healing.

Hidrogéis compreendem uma importante classe de materiais poliméricos adequados à aplicação como curativos de feridas e queimaduras. A estrutura tridimensional hidrofílica dos hidrogéis permite que estes mantenham a umidade ideal no leito das feridas, absorvam o exsudato e não causem danos ao novo tecido durante as trocas dos curativos. No caso dos hidrogéis, essas trocas podem ser menos frequentes. Além disso, curativos que auxiliem na remoção de tecidos necrosados e ainda sejam capazes de oferecer tratamentos extras que acelerem o processo de cicatrização são desejáveis. Este trabalho apresenta a produção de materiais à base de hidrogel capazes de auxiliar neste processo de diferentes maneiras. Primeiramente, são apresentados hidrogéis formados a partir de nanofibras de poli(N-vinil-2-pirrolidona) (PVP) produzidas por eletrofiação, seguido da reticulação através da utilização de radiação UV-C ou reação de Fenton. A utilização da eletrofiação como técnica auxiliar na formação dos hidrogéis permitiu o controle da porosidade através da formação de fibras de diferentes diâmetros. A evidência de tal propriedade foi constatada através da produção de materiais que apresentam diferentes perfis de liberação da proteína modelo albumina de soro bovino (BSA). O hidrogel de PVP nanoestruturado foi capaz de liberar e manter a atividade da colagenase, uma importante enzima aplicada no tratamento de feridas via desbridamento enzimático, durante as 48 horas em que foi avaliado. Além disso, hidrogéis bactericidas nanoestruturados foram produzidos a partir de nanocompósitos de PVP e nanopartículas de prata (AgNP) produzidos por eletrofiação. Esses hidrogéis apresentaram propriedades térmicas semelhantes aos hidrogéis sem AgNP, diminuindo, contudo, a sua capacidade de intumescimento. Esses hidrogéis mostraram-se ativos contra bactérias gram-positivas e gram-negativas a partir de 100 ppm de AgNPs. Adicionalmente, foi estudada a formação de um hidrogel modelo composto PVP/AgNP/Imidazol, almejando-se a produção de um material bactericida-fungicida a base de hidrogel. Este hidrogel apresentou atividade conta três espécies de Candida a partir de 500 ppm de imidazol no material. Embora exista a formação de um complexo estável entre AgNP e Imidazol, cálculos teóricos e a constatação da atividade fungicida corroboram com o fato de que derivados imidazólicos podem ser liberados a partir deste hidrogel híbrido. A produção de hidrogéis físicos compostos por blendas de PVP/Polianidridos sintetizados a partir de derivados de hidroxicinamatos e ácido salicílico, capazes de liberar moléculas de interesse biológico quando parcialmente degradados hidroliticamente, também é descrita neste trabalho. Os resultados indicam que interações hidrofóbicas entre a PVP e os polianidridos sintetizados podem ser responsáveis pela formação dos hidrogéis físicos e pela miscibilidade das blendas produzidas. Os hidrogéis físicos de PVP/Polianidridos foram obtidos na forma de filmes por evaporação do solvente. Micro- e nanofibras também foram obtidas por eletrofiação. Desta maneira, o presente trabalho contribui com o desenvolvimento de uma geração de curativos multifuncionais aplicados no tratamento de feridas crônicas e queimaduras. / Hydrogels comprise an important class of polymeric materials that finds application as wound and burn dressings. The hydrophilic three-dimensional structure of hydrogels helps to provide the ideal humidity at the wound bed, to remove exsudates and to prevent damages to the new tissue during dressing substitution. Furthermore, these wound dressings are able to remove necrotic tissues and, therefore, capable to offer extra treatments that would benefit the healing processes. This work describes the production of hydrogel based materials that are able to act in wound healing by different ways. First, it is presented hydrogels composed of poly(N-vinyl-2-pyrrolidone) (PVP) nanofibers produced by electrospinning, followed by its crosslinking using UV-C radiation or Fenton reaction. The use of electrospinning in the hydrogel formation allowed porosity control by obtaining fibers of different diameters. This was evidenced by achieving materials that present different release profiles of the model protein bovine serum albumin (BSA). The nanostructured PVP hydrogel was capable of releasing and maintaining collagenase activity during 48 hour of evaluation. This is an important enzyme that find application in wound healing based on enzymatic debridement. Moreover, nanostructured bactericidal hydrogels were produced from PVP and silver nanoparticles (AgNP) composite through electrospinning, resulting in hydrogels with thermal properties similar to those hydrogels without AgNP, decreasing its swelling ability. These hydrogels were active against gram-positives and gram-negatives bacteria starting from 100 ppm of AgNP. In addition, the production of a model hydrogel composed by PVP/AgNP/Imidazole was investigated, aiming at a bactericidal-fungicidal hydrogel based material. This hydrogel was active against three Candida having 500 ppm of imidazole into the structure. In spite of the formation of a stable complex between AgNP and imidazole, theoretic calculations and the observed fungicidal activity corroborate with the fact that imidazoles derivatives can be released from this hybrid hydrogel. Physical hydrogels composed of PVP/Polyanhydrides blends were synthesized from hydroxycinammates derivatives and salicylic acid. These materials which were capable of releasing molecules with biological potential upon hydrolysis, are also described in this work. The results indicate that hydrophobic interactions between PVP and the synthesized polyanhydrides could be responsible for the hydrogel formation and blend miscibility as well. PVP/Polyanhydride physical hydrogels were obtained from cast films. Micro- and nanofibers were also obtained by electrospinning. Thus, the present work contributes with the development of the new generation of smart dressings for wound and burn healing.

Regeneration, a spontaneous response of bones in response to injuries, infections and fractures, is severely compromised in certain clinical circumstances. Unfortunately, several shortcomings are associated with the current treatment of bone grafting method such as donor shortage and immune response for allografts and donor morbidity for autografts. Thus, the development of clinical alternates is essential. One promising adjunct method is bone tissue engineering that includes the implantation of a scaffold containing the cells with the supplementation of suitable growth factors. Among the various classes of materials, biodegradable polymers are commonly preferred because their use does not necessitate a secondary surgery for their removal after the intended use. Commercially available polymers such as poly (lactic- co- glycolic acid) and polycaprolactone are expensive and degrade slowly. This motivates the development of novel synthetic biodegradable polymers that are affordable and can be tuned to tailor for specific biomedical applications. The primary aim of this thesis is to synthesize effective biodegradable polymers for drug delivery and bone tissue engineering. The properties of these polymers such as modulus, hydrophobicity and crosslinking etc. were tailored based on the variations in chemical bonds, chain lengths and the molar stoichiometric ratios of the monomers for specific clinical applications. Based on the above variations, degradation and release kinetics were tuned. The cytocompatibilty properties for these polymers were studied and suitable mineralization studies were conducted to determine their potential for bone regeneration.