Nanoparticle usage continues to increase in everyday products, from cosmetics to food preservation coatings, drug delivery to polymer fillers. Their characterization and synthesis is of utmost importance to ensure safety and improved product quality. Nanoparticles can be sourced naturally or synthetically fabricated. Cellulose nanocrystals (CNCs) are rod-like nanoparticles that can be isolated from nature. Reliable methods of characterization are necessary to ensure quality control. However, their physical characteristics cause challenges for imaging under transmission electron microscopy (TEM) with a high enough resolution for dimensional analysis. Heavy metal staining such as radioactive uranyl acetate is often used to increase contrast and TEM sample substrate preparation techniques often use expensive equipment such as glow discharge in order to prevent CNC agglomeration. A method to reliably produce TEM images of CNCs without using radioactive stains or expensive glow discharge equipment was developed, using a vanadium-based stain branded NanoVan® and bovine serum albumin to keep CNCs dispersed while drying on the TEM substrate. Due to their aspect ratio, there is also concern of toxicity to the lungs. The concentration of CNCs in air in production facilities must be monitored, but there is currently no method tailored to CNCs. A method using UV-vis spectroscopy, dynamic light scattering, TEM, and scanning mobility particle sizer in conjunction with impinger collectors was developed for monitoring aerosolized CNC concentration. Synthetic nanoparticles are often used for controlled drug delivery systems. A new peptide drug termed αCT1 has been shown to interact with cell communication in a way that promotes wound healing, reduces inflammation and scarring, and aids in cancer therapy. However, the peptide's half-life in the body is estimated to be less than a day, which is not conducive to long-term treatments. Controlling its release into the body over several weeks can decrease the number of doses required, which is especially useful for glioblastoma treatment. Poly(lactic-co-glycolic acid) (PLGA) is often used for drug encapsulation since it hydrolyzes in the body and is biocompatible. Two methods of αCT1 encapsulation in PLGA were explored. It was found that flash nanoprecipitation increased loading of αCT1 in the particles by 1-2 orders of magnitude compared with the double emulsion method. Particles released αCT1 over three weeks and were non-cytotoxic. / PHD / Nanoparticle usage continues to increase in everyday products, from cosmetics to food preservation coatings, drug delivery to polymer fillers. Understanding the nature of nanoparticles is important to ensure safety and quality of commercial products, and production of particles allows for tailoring for specific applications. In this work, a technique to more easily create samples of cellulose nanocrystals (CNCs) for electron microscopy is developed. Electron microscopy can then be used to measure the size of these rod-like particles. Then, the technique is used to help develop a method to measure the concentration of CNCs in air. CNCs may irritate the lungs, so development of a way to measure their concentration in air is important to ensure safety of plant workers and consumers of CNCs. Characterization techniques of CNCs were used for synthesized particles used for brain cancer treatment. Synthesized particles contain the drug αCT1, which has been shown to reduce glioblastoma, or brain cancer, from becoming resistant to chemotherapy. These particles were made using poly(lactic-co-glycolic acid) (PLGA), a polymer that degrades in the body into lactic acid and glycolic acid. PLGA particles released αCT1 over three weeks and are of a size that is compatible with the brain. However, loading of the drug was low when using the first synthesis method. By switching particle synthesis methods, drug loading in the particles was increased by 1-2 orders of magnitude.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

Introdução: A aplicação clínica de biomateriais está se expandindo para diversas especialidades médicas. Dentre os diversos tipos de biomateriais, os classificados como temporários merecem atenção especial, pois são assimilados pelo organismo após exercerem sua função, evitando, assim, procedimento cirúrgico para sua retirada. O copolímero de poli (ácido láctico-co-glicólico) (PLGA) é um tipo de biomaterial temporário, rotineiramente utilizado na medicina na forma de fios de sutura e implantes ortopédicos. A mistura do PLGA com o poli (isopreno) resulta em uma blenda (PLGA / PI), de alta resistência e tenacidade, que foi desenvolvida pelo Laboratório de Biomateriais do Instituto de Engenharia da UFRGS. Entretanto, não existem estudos “in vivo” testando a reação óssea desta blenda. Objetivo: Testar a histotoxicidade da blenda de PLGA / PI em relação ao biopolímero já consagrado PLGA. Método: Foram utilizados 46 ratos machos wistar (Rattusnorvegicus - linhagem albina), divididos em 2 grupos conforme o material implantado (PLGA ou PLGA / PI) na calota craniana, e subdivididos em tempos de morte (15, 30, 60 e 90 dias). Os procedimentos foram realizados na Unidade de Experimentação Animal (UEA) do Hospital de Clínicas de Porto Alegre (HCPA). Após a morte, a calota craniana foi retirada, submetida ao exame histopatológico e aplicado o escore de Dadas e cols (14) modificado. Resultados: A diferença da histotoxicidade dos dois materiais não foi significativa nos períodos 15, 30 e 90 dias, porém foi significativa no período 60 dias. Conclusão: A histotoxicidade do PLGA / PI, ao final do estudo (90 dias), foi semelhante ao PLGA, demonstrando equivalência em longo prazo. O período de 60 dias pós-cirúrgico (grupo da blenda) foi o único em que a histotoxicidade mostrou-se significativamente maior. Mais estudos devem ser feitos para melhorar o entendimento desta variação. / Introduction: Clinical application of biomaterials is expanding to various medical specialties. Among the different types of biomaterials, those classified as temporary deserve special attention because they are assimilated by the body after exercising their function, thereby avoiding surgical procedure for their removal. Co-polymer poly (lactic-co-glycolic acid) (PLGA) is a type of temporary biomaterial, routinely used in medicine as suture threads and orthopedic implants. The mixture of PLGA with poly (isoprene) results in a high-strength and thoughness blend (PLGA / PI), developed by the Biomaterials Laboratory of the Engineering Institute/ UFRGS. However, there are no studies “in vivo” testing the bone reaction of that blend. Objective: To Test histotoxicity of PLGA / PI blend over the already established biopolymer, PLGA. Method: Forty six male Wistar rats (Rattus norvegicus – albino strain), divided into 2 groups according to the material (PLGA or PLGA / PI) implanted in the skull and sub divided into periods of death (15, 30, 60 and 90 days). The procedures were developed in the Animal Experiment Unit (AEU) of Hospital de Clínicas de Porto Alegre (HCPA). After death, the skull was removed, submitted to histopathologic examination and the modified Dadas’ et all score was used (14). Results: The histotoxicity difference of the two materials was not significant in the periods of 15, 30 and 90 days, but it was significant in the period of 60 days. Conclusion: At the end of the study (90 days), the PLGA / PI histotoxicity was similar to PLGA, showing longterm equivalence. The 60-day post-surgical period was the only one in which histotoxicity was significantly higher (blend group). More studies shall be done in in order to better understand that variation.

MPharm. Faculty of Health Sciences, University of the Witwatersrand, 2007

The biomaterial component of a tissue engineered device has been shown to enhance the immune response to a co-delivered model shed antigen. The purpose of this research was to investigate in vivo the differential level of the immune response toward different forms of the biomaterial. A model shed antigen, ovalbumin (OVA), was incorporated into polymeric biomaterial carriers made of 50:50 poly(lactic-co-glycolic acid) (PLGA) in the form of microparticles (MP) or scaffolds (SC). These MP and SC biomaterial carrier vehicles with incorporated antigen were then injected or implanted, respectively, into C57BL6 mice to investigate the differential level of the immune response towards OVA controlled release from PLGA MP and PLGA SC. For each polymeric carrier, the resulting time-dependent systemic humoral immune response towards the incorporated OVA, the OVA-specific IgG concentration and isotypes (IgG2a or IgG1, indicating a predominant Th1 or Th2 response, respectively) were determined using ELISA. To assess the differential level of the immune response depending on the form of PLGA, the total amounts of polymer and OVA delivered were kept constant as well as the release rate of OVA. The in vitro protein release kinetics were studied for both PLGA MPs and PLGA scaffolds to examine the release rate of OVA from the polymeric carriers. The level of the humoral immune response was higher and sustained for OVA released from PLGA SC which were implanted with associated tissue damage, and lower and transient when the same amount of polymer and OVA were delivered from PLGA MP, which were minimally invasively delivered by injection. This immune response was primarily Th2 helper T cell-dependent as exemplified by the predominance of IgG1 isotype, although for the strong adjuvant, Complete Freunds adjuvant (CFA), and PLGA SC carriers the anti-OVA IgG2a isotype levels were also significant, potentially indicating both a Th2 and Th1 response. The PLGA SC and PLGA MP exhibited similar protein release kinetics, releasing similar amounts of OVA at each time point. Each carrier incubated contained the same ratio of OVA to polymer. In vitro protein release kinetics experiments suggest that the rate of release of OVA from PLGA SC and PLGA MP was similar, and therefore the enhanced immune response induced by PLGA SC is most likely due to danger signals from implantation which primed the system for an enhanced immune response and not from a difference in concentration of OVA released from the carriers.

PLGA have already been successfully applied for controlled drug delivery systems by the pharmaceutical industry due to its biocompatibility, biodegradability and ease of processing. It has recently further been developed and formulated into a form of nanoparticle. The single emulsion evaporation method was used to prepare nanoparticles in this study. By varying different parameters such as the concentration of regents, the type of surfactant and emulsion method, different particle sizes and size distribution of PLGA nanoparticles could be obtained. The stability of PLGA nanoparticles was further investigated by assessing their thermal property over a certain period of time using DSC. The decrease of Tg confirmed the hydration and degradation of PLGA polymers and nanoparticles. The changes of surface morphology showed that the nanoparticles were in spherical shape and maintained smooth surface before the storage, whereas they started to lose their original shapes as well as agglomerate to each other after 2-week storage. These results suggested that there was an erosion and degradation of PLGA nanoparticles during storage. Ibuprofen-loaded PLGA nanoparticles have been successfully prepared by o/w single emulsion evaporation method. During the stability study, a faster degradation rate compared to non-loaded PLGA nanoparticles was exhibited, showing that Ibuprofen increased the degradation rate of PLGA nanoparticles. According to the results of drug releasing study, PLGA nanoparticles exhibiting a slower drug release rate than pure drug which proved that drug-nanoparticule system could effectively increase the stability of drugs. PLGA polymer is a potential material for drug delivery system.

Introduction: Bone defects and non-unions caused by trauma, tumor resection, pathological degeneration, or congenital deformity pose a great challenge in the field of orthopedics. Traditionally, these defects have been repaired by using autografts and allografts. Autografts have set the gold standard for clinical bone repair because of their osteoconductivity, osteoinductivity and osteogenicity. Nevertheless, the application of autografts is limited because of donor availability and donor site morbidity. Allografts have the advantage that the tissues are readily available and can be easily applied, especially when large segments of bone are to be reconstructed. However, their use is also limited by the risk of disease transfer and immune rejection. To circumvent these limitations tissue engineering has evolved as a means to develop viable bone grafts. An ideal bone graft should be both osteoconductive and osteoinductive, biomechanically strong, minimally antigenic, and eliminates donor site morbidity and quantity issues. The biodegradable polymer, Poly lactic-co-glycolic acid (PLAGA) was chosen because of its commercial availability, biocompatibility, non-immunogenicity, controlled degradation rate, and its ability to promote optimal cell growth. To improve the mechanical properties of PLAGA, Single Walled Carbon Nanotubes (SWCNT) were used as a reinforcing material to fabricate composite scaffolds. The overall goal of this project is to develop a Single Walled Carbon Nanotube composite (SWCNT/PLAGA) for bone regeneration and to examine the interaction of MC3T3-E1 cells (mouse fibroblasts) and hBMSCs (human bone marrow derived stem cells) with the SWCNT/PLAGA composite via focusing on extracellular matrix production and mineralization; and to evaluate its toxicity and bio-compatibility in-vivo in a rat subcutaneous implant model. We hypothesize that reinforcement of PLAGA with SWCNT to fabricate SWCNT/PLAGA composites increases both the mechanical strength of the composites as well as the cell proliferation rate on the surface of the composites while expressing osteoblasts phenotypic, differentiation and mineralization markers; and SWCNT/PLAGA composites are biocompatible and non-toxic, and are ideal candidates for bone tissue engineering. Methods: PLAGA and SWCNT/PLAGA composites were fabricated with various amounts of SWCNT (5, 10, 20, 40 and 100mg), characterized and degradation studies were performed. PLAGA (poly lactic-co-glycolic acid) and SWCNT/PLAGA microspheres and composites were fabricated; characterized and mechanical testing was performed. Cells were seeded and cell adhesion/morphology, growth/survival, proliferation and gene expression analysis were performed to evaluate biocompatibility. Sprague-Dawley rats were implanted subcutaneously with Sham, poly lactic-co-glycolic acid (PLAGA) and SWCNT/PLAGA composites, and sacrificed at 2, 4, 8 and 12 week post-implantation. The animals were observed for signs of morbidity, overt toxicity, weight gain, food consumption, hematological and urinalysis parameters, and histopathology. Results: Imaging studies demonstrated uniform incorporation of SWCNT into the PLAGA matrix and addition of SWCNT did not affect the degradation rate. Composites with 10mg SWCNT resulted in highest rate of cell proliferation (p<0.05) among all composites. Imaging studies demonstrated microspheres with uniform shape and smooth surfaces, and uniform incorporation of SWCNT into PLAGA matrix. The microspheres bonded in a random packing manner while maintaining spacing, thus resembling trabeculae of cancellous bone. Addition of 10mg SWCNT led to greater compressive modulus and ultimate compressive strength. Imaging studies revealed that MC3T3-E1 cells adhered, grew/survived, and exhibited normal, non-stressed morphology on the composites. SWCNT/PLAGA composites exhibited higher cell proliferation rate and gene expression compared to PLAGA. No mortality and clinical signs were observed. All the groups showed consistent weight gain and rate-of-gain for each group was similar. All the groups exhibited similar pattern for food consumption. No difference in urinalysis parameters, hematological parameters; and absolute and relative organ weight was observed. A mild to moderate summary toxicity (sumtox) score was observed for animals treated with the PLAGA and SWCNT/PLAGA whereas the sham animals did not show any response. At all the time intervals both PLAGA and SWCNT/PLAGA showed a significantly higher sumtox score compared to the Sham group. However, there was no significant difference between PLAGA and SWCNT/PLAGA groups. Conclusion: Our SWCNT/PLAGA composites, which possess high mechanical strength and mimic the microstructure of human trabecular bone, displayed tissue compatibility similar to PLAGA, a well known biocompatible polymer over the 12 week study. Thus, the results obtained demonstrate the potential of SWCNT/PLAGA composites for application in BTE and musculoskeletal regeneration. Future studies will be designed to evaluate the efficacy of SWCNT/PLAGA composites in bone regeneration in a non-union ulnar bone defect rabbit model. As interest in carbon nanotube technology increases, studies must be performed to fully evaluate these novel materials at a nonclinical level to assess their safety. The ability to produce composites capable of promoting bone growth will have a significant impact on tissue regeneration and will allow greater functional recovery in injured patients.

Tobacco smoking has consistently been the leading cause of preventable diseases and premature deaths. Currently, pharmacological interventions have only shown limited smoking cessation efficacy and sometimes are associated with severe side effects. As an alternative, nicotine vaccines have emerged as a promising strategy to combating nicotine addiction. However, conventional conjugate nicotine vaccines have shown limited ability to induce a sufficiently strong immune response due to their intrinsic shortfalls. In this study, a lipid-poly(lactic-co-glycolic acid) (PLGA) nanoparticle-based next-generation nicotine vaccine has been developed to overcome the drawbacks of conjugate nicotine vaccines. Also, the influence of multiple factors, including nanoparticle size, hapten density, hapten localization, carrier protein, and molecular adjuvants, on its immunogenicity has been investigated. Results indicated that all these studied factors significantly affected the immunological efficacy of the nicotine nanovaccine. First, 100 nm nanovaccine was found to elicit a significantly higher anti-nicotine antibody titer than the 500 nm nanovaccine. Secondly, the high-density nanovaccine exhibited a better immunological efficacy than the low- and medium-density counterparts. Thirdly, the nanovaccine with hapten localized on both carrier protein and nanoparticle surface induced a significantly higher anti-nicotine antibody titer and had a considerably better ability to block nicotine from entering the brain of mice than the nanovaccines with hapten localized only on carrier protein or nanoparticle surface. Fourthly, the nanovaccines carrying cross reactive materials 197 (CRM197) or tetanus toxoid (TT) showed a better immunological efficacy than the nanovaccines using keyhole limpet hemocyanin (KLH) or KLH subunit as carrier proteins. Finally, the co-delivery of monophosphoryl lipid A (MPLA) and Resiquimod (R848) achieved a considerably higher antibody titer and brain nicotine reduction than only using MPLA or R848 alone as adjuvants. Collectively, the findings from this study may lead to a better understanding of the impact of multiple factors on the immunological efficacy of the hybrid nanoparticle-based nicotine nanovaccine. The findings may also provide significant guidance for the development of other drug abuse and nanoparticle-based vaccines. In addition, the optimized lipid-PLGA hybrid nanoparticle-based nicotine nanovaccine obtained by modulating the studied factors can be a promising candidate as the next-generation nicotine vaccine for treating nicotine addiction. / PHD / Tobacco smoking is prevalent and represents one of the largest public health concerns in the world. Tobacco use remains to be the leading cause of preventable diseases and premature deaths. Typically, smokers without medications experience huge difficulty in quitting smoking due to the addictiveness of nicotine. Although several pharmacological interventions are available to smokers, they have only shown limited smoking cessation efficacy. In recent years, nicotine vaccines that can induce the production of nicotine specific antibodies have been proposed as a promising strategy for smoking cessation. Currently, almost all the already-developed nicotine vaccines are conjugate nicotine vaccines in which nicotine haptens are attached to a carrier protein for presentation. However, conventional conjugate nicotine vaccines suffer from many innate shortcomings that largely limit their immunological efficacy. This study aimed to develop a nanoparticle-based next-generation nicotine vaccine to overcome the drawbacks of conventional conjugate nicotine vaccines. Because many factors may potentially affect the immunological efficacy of a nicotine vaccine, this study focused on investigating the influence of multiple factors, including nanoparticle size, hapten density, hapten localization, carrier protein, and molecular adjuvants, on the immunological efficacy of the developed hybrid nanoparticle-based nicotine vaccine. Results from this study revealed that all these studied factors significantly influenced the immunological efficacy of the nicotine nanovaccine. By modulating v these factors, enhanced immune responses that can result in higher titers of anti-nicotine antibodies and better ability in blocking nicotine from entering the brain of mice could be achieved. The findings from this study would lead to a better understanding of how multiple factors influence the immunological efficacy of a nanoparticle-based nicotine vaccine. In addition, the findings from this study can also provide significant guidance for the development of other drug abuse and nanoparticle-based vaccines. More importantly, the hybrid nanoparticle-based nicotine nanovaccine can be a promising candidate for smoking cessation.

Capsaicin, prostaglandin E2 (PGE2) and polydopamine (PDA) were used to target macrophage and myofibroblast activity to reduce biomaterial-induced fibrosis. The lifetime and efficacy of implantable biomedical devices are determined by the foreign body response. Immediately after implantation, proteins nonspecifically adsorb onto the material and initiate inflammation. Macrophages recruited to the site can differentiate into M1 and M2 phenotypes and upregulate inflammation and fibrosis which interferes with the intended function. M1 macrophages secrete pro-inflammatory mediators that induce chronic inflammation and promote myofibroblast differentiation while M2 macrophages are wound healing cells that suppress inflammation and regulate fibroblast activity. The fibrotic tissue is developed by myofibroblasts which produce collagen in an unregulated fashion. Collagen thickening and biomaterial encapsulation decreases efficacy and sensitive of biomedical devices. We investigated the in vitro and in vivo effects of capsaicin, PGE2 and polydopamine surface modification on macrophages and myofibroblasts. Capsaicin and PGE2 reduced poly(lactic-co-glycolic) acid (PLGA)-induced fibrosis by promoting M2 macrophage phenotype to secrete anti-inflammatory IL-10 and suppressing myofibroblast marker α-smooth muscle actin (α-SMA). Capsaicin decreased collagen by 40% and upregulated IL-10 secretion by 35% while PGE2 reduced collagen by 55% after 14 days of implantation and 40% less collagen after 42 days. PDA was used to bind an anti-fibrotic compound to the surface of a poly(dimethyl siloxane) (PDMS-PDA) to reduce fibrosis. However, PDMS-PDA controls gave an unexpected result by reducing fibrosis to the same extent as anti-fibrotic compound bound PDMS- v PDA. PDA modification reduced cellularity by 50% and significantly decreased collagen thickness by 30%. Overall, our results showed that biomaterial-induced fibrosis can be reduced by promoting M2 macrophage activity and inhibiting myofibroblast differentiation. This research demonstrates three compounds that have potential to reduce fibrosis and extend the lifetime and efficacy of implantable biomedical devices. / Thesis / Master of Applied Science (MASc) / Capsaicin, prostaglandin E2 (PGE2) and polydopamine were used to reduce scar tissue development around implanted polymers. Biomedical devices implanted in the body can undergo severe scar tissue formation, or fibrosis, and fail. Fibrosis is described by the accumulation of collagen and encapsulation of an implanted polymer. Macrophages regulate fibrosis by secreting pro-fibrotic compounds and myofibroblasts produce unregulated amounts of collagen. In this thesis, capsaicin, PGE2 and polydopamine were incorporated into implants to target macrophage and myofibroblast activity and reduce fibrosis in mice. Capsaicin and PGE2, released from a degradable polymer, altered macrophages to secrete anti-fibrotic compounds and decreased collagen by 40% and 55%, respectively. Polydopamine surface modified implants gave an unexpected result and suppressed overall cell activity to reduce fibrosis by 30%. The research conducted shows the potential of these compounds to reduce fibrosis and extend the lifetime of implantable devices.