This work has explored the capability of a mixed film composed of oligonucleotides and oligomers to improve the selectivity for the detection of fully complementary oligonucleotide targets in comparison to partially complementary targets which have one and three base-pair mismatched sites. The intention was to introduce a “matrix isolation” effect on oligonucleotide probe molecules by surrounding the probes with oligomers, thereby reducing oligonucleotide-to-oligonucleotide and/or oligonucleotide-to-surface interactions. This resulted in a more homogeneous environment for probes, thereby minimizing the dispersity of energetics associated with formation of double-stranded hybrids. The mixed film was constructed by immobilizing pre-synthesized oligonucleotides onto a mixed aminosilane layer and then growing the oligomer portion by surface-initiated atom transfer radical polymerization (ATRP) of 2-hydroxy methacrylate (PHEMA). The performance of the mixed film was compared to films composed of only oligonucleotides in a series of hybridization and melt curve experiments. Surface characterization techniques were used to confirm the growth of the oligomer portion as well as the presence of both oligonucleotides and oligomer components. Polyatomic bismuth cluster ions as sources for time-of-flight secondary ion mass spectrometry experiments could detect both components of the mixed film at a high sensitivity even though the oligomer portion was at least 200-fold in excess. At the various ionic strengths investigated, the mixed films were found to increase the selectivity for fully complementary targets over mismatched targets by increasing the sharpness of melt curves and melting temperature differences (delta Tm) by 2- to 3-fold, and by reducing non-specific adsorption. This resulted in improved resolution between the melt curves of fully and partially complementary targets. A fluorescence lifetime investigation of the Cy3 emission demonstrated that Cy3-labeled oligonucleotide probes experienced a more rigid microenvironment in the mixed films. These experiments demonstrated that a mixed film composed of oligonucleotides and PHEMA can be prepared on silica-based substrates, and that they can improve the selectivity for SNP discrimination compared to conventional oligonucleotide films.

O objetivo deste estudo é caracterizar a influência da relação dos componentes hidrófilos de uma resina adesiva experimental em suas propriedades intrínsecas. Para isso, foram adicionados a um sistema polimérico (BisGMA, BisEMA e TEGDMA), diferentes concentrações de HEMA totalizando quatro grupos: 0%, 15%, 30% e 50% de HEMA. As misturas foram caracterizadas quanto à cinética de polimerização através de Fourier Transformed Infrared (FTIR) em tempo real e a taxa de polimerização (RP) foi calculada através da análise de regressão não-linear; resistência coesiva à tração utilizando-se uma máquina de ensaios mecânicos com corpos de prova em forma de ampulheta com área de constrição de 1mm2; e, sorção e solubilidade baseada na normatização ISO 4049. Houve correlação entre o tempo de fotoativação e o grau de conversão para todos os grupos (r2>0.98). O grupo 0% apresentou RPMÁX elevada nos segundos iniciais de fotoativação com, no entanto, uma brusca redução. As adições de 30% e 50% de HEMA determinaram uma RPMÁX do copolímero menor que em 15% e 0%. Não houve diferença entre o grau de conversão final (60s) entre os grupos de 0, 15 e 30%, sendo 59,7%, 61,4% e 55,2% respectivamente. As adições de 30% e 50% de HEMA determinaram a redução da resistência coesiva dos polímeros (p<0,05) quando comparado aos grupos 0% e 15%. Grupos com maiores adições de HEMA apresentaram maiores graus de sorção e solubilidade (p<0,05). Maiores adições de HEMA resultaram em polímeros mais propensos à degradação. / The aim of this study was to characterize the influence of hydrophilic monomers ratios on an experimental adhesive resin properties. Four groups, with 0, 15, 30 and 50% (wt) addiction of HEMA were prepared. Comonomer blends (BisGMA, BisEMA, TEGDMA and HEMA) were analyzed with kinetics of polimerization real time FTIR and the rate of polymerization was calculated by non-linear regression analysis. Ultimate tensile strength was tested using a universal testing machine (Emic) with hour-glass shaped specimens with cross-sectional area 1mm2. Water sorption and solubility tests were based on ISO 4049. For All groups was a positive and strong correlation between light-activation time and degree of conversion (r2>0,98). Group 0% of HEMA showed higher RPMÁX in early seconds with a suddenly decrease. Additions of 30% and 50% of HEMA determined a RPMÁX lower than the comonomer blends with 15% and 0% of HEMA. There was no statistical difference of final degree of conversion of the tested groups, varying from 63% to 67% . The additions of 30% and 50% of HEMA determined reduction of ultimate tensile strength of the polymers formed (p<0,05) when compared to 0% and 15 % of HEMA. Groups with higher additions of HEMA showed higher water sorption and solubility (p<0,05). Higher additions of HEMA resulted in polymers more prone to degradation.

O objetivo deste estudo foi sintetizar e caracterizar monômeros metacrilamidas, desenvolver, caracterizar e avaliar as propriedades de sistemas adesivo convencional de três passos. Foram sintetizadas bis(metacrilamida)s e tris(metacrilamida) e caracterizadas por espectroscopia de Infravermelho por Transformada de Fourier (FTIR), Ressonância Magnética Nuclear (RMN) de 1H e 13C, por Cromatografia Líquida de Alta Eficiência com Espectrometria de Massas (UHPLC-QTOF-MS) e Calorimetria Exploratória Diferencial Modulada (MDSC). Quatro bis(metacrilamidas), (1) N,N’-(propane-1,3-diyl)bis(N-ethyl-2-methylacrylamide), (2) N,N’-(octane-1,8-diyl)bis(2-methylacrylamide), (3) N,N’-(butane-1,4-diyl)bis(2-methacrylamide) e (4) N,N’-(1,4 phenylene)bis(2-methylacrylamide)monômero (1), (2), (3) e (4), e uma tris(metacrilamida) TMA, foram sintetizadas. Pela análise de FTIR-ATR foram observadas as bandas correspondentes ao estiramento do grupo C=O (1660 cm-1), C=C (1610 cm-1), N-H (3300 cm-1) e C-N (1520 cm-1). As análises de RMN identificaram a presença das ligações duplas referentes aos grupos metacrilamidas em deslocamentos químicos entre 5,3 e 5,8 ppm para 1H e entre 120 e 140 ppm para 13C.Os valores de massa exata m/z foram: 267,2068, 281,2222, 225,1595, 245,1283 e 351,2385 g/mol para os monômeros (1), (2), (3), (4) e TMA, respectivamente. A cinética de polimerização do TMA e dos adesivos experimentais contendo 2-hidroxietil acrilamida (HEAA) ou 2-hidroxietil metacrilato (HEMA) com as seguintes formulações foram investigadas por meio de DSC-PCA, n=3: TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%, TMA50%/HEMA50%, BisGMA/HEAA/TMA e BisGMA/HEMA.Características e propriedades mecânicas das resinas adesivas BisGMA/HEAA/TMA e BisGMA/HEMA foram avaliadas por resistência coesiva (UTS, n=5), degradação em solvente (ΔKHN, n=5), ângulo de contato (n=5), microtração (μTBS, n=20) e análise de fratura. Um primer a base de acrilamidas foi desenvolvido (H2O/HEAA/AMPS) (2-acrylamida-2-methilpropano ácido sulfônico) para ser utilizado no grupo experimental com metacrilamidas. Os valores de pH e ângulo de contato do primer experimental foram comparados com o primer do ScotchBond Multi-purpose (grupo controle). O monômero (1) resultou em um monômero amarelo claro de baixa viscosidade, entretanto, não apresentou foto ou termopolimerização. A energia de ativação determinada pelo método de Kissinger foi – 165,8 kJmol-1; -182,7 kJmol-1 e -156,7 kJmol-1 para os monômeros (2), (3) e (4), respectivamente. Sistemas adesivos convencionais de três passos a base de metacrilamidas e a base de metacrilatos foram desenvolvidos. Resinas adesivas contendo somente HEAA e TMA (TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%) apresentaram grau de conversão abaixo de 40% após 40 s de fotoativação. Alto grau de conversão (acima de 60%) só foi encontrado para as resinas adesivas BisGMA/HEAA/TMA e BisGMA/HEMA e sem diferença significativa entre elas, p>0,05. Os valores de UTS (BisGMA/HEMA- 67,7 ±5 MPa e BisGMA/HEAA/TMA- 60,5 ±7 MPa), μTBS (BisGMA/HEMA- 57 ± 14 MPa e BisGMA/HEAA/TMA- 53,1 ±15 MPa) e ângulo de contato (BisGMA/HEMA- 39,5 ±9 e BisGMA/HEAA/TMA- 46,7 ±15) não apresentaram diferença estatística, p>0.05. O primer experimental apresentou um valor pH mais baixo (2,7) bem como de ângulo de contato (18,5 ±5) em relação ao comercial (pH-4 e θ-33,5 ±4). A síntese proposta para os monômeros (1), (2), (3), (4) e TMA foi caracterizada nesse trabalho. Um primer somente com acrilamidas foi desenvolvido e a presença do novo monômero TMA na resina adesiva BisGMA/HEAA permitiu a formulação de um sistema adesivo convencional de três passos sem a presença do monômero HEMA. / The aim of this study was synthesized and characterizes methacrylamides monomers, development, characterizer and evaluated the properties of 3-step etch-and-rise adhesive system. Bis(methacrylamide)s and tris(methacrylamide) were synthesized. The monomer structures were confirmed by 1H and 13C Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FTIR-ATR), Ultra-high liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) and modulated differential scanning calorimetry (mDSC). Four bis(methacrylamide)s monomers (1) N,N’-(propane-1,3-diyl)bis(N-ethyl-2-methylacrylamide), (2) N,N’-(octane-1,8-diyl)bis(2-methylacrylamide), (3) N,N’-(butane-1,4-diyl)bis(2-methacrylamide) and (4) N,N’-(1,4 phenylene)bis(2-methylacrylamide) and one tris(methacrylamide), TMA, were synthesized. All IR spectra of the monomers showed the C=C axial deformation at 1610 cm-1. The 1H NMR spectra the olefinic hydrogens were observed at 5.3 an 5.8 ppm and in the 13C NMR, the vinylic carbons at 120 and 140 ppm. The exact m/z values were: 267.2068, 281.2222, 225.1595, 245.1283 and 351.2385 g/mol for monomers (1), (2), (3), (4) and TMA respectively. Monomer (1) not presented photo (DSC-PCA) or thermal polymerization. The activation energy determined using Kissinger methodology was: - 165.8 kJmol-1; -182.7 kJmol-1 and -156.7 kJmol-1 for monomers (2), (3) and (4), respectively. 3-step adhesive systems with methacrylamides and methcrylates were development. Kinetics of photopolymerization of TMA and experimental adhesive resin containing 2-hydroxyethylacrylamide (HEAA) or 2-hydroxyethylmethacrylate (HEMA) in the following formulations: (TMA 33%/HEAA 66%, TMA 50%/HEAA 50%, TMA66%/HEAA33%, TMA50%/HEMA50%, BisGMA66%/HEAA24%/TMA10% and BisGMA66%/HEMA33%) were investigated through DSC-PCA. Characteristics and mechanical properties for BisGMA 66%/HEAA 24%/TMA 10% and BisGMA 66%/HEMA 33% adhesives were evaluated with ultimate tensile strength (UTS, n=5), softening in solvent (ΔKHN, n=5), contact angle (n=5), microtensile bond strength (μTBS, n=20) and failure analysis. A primer was also formulated with H2O/HEAA/AMPS (2-acrylamido-2-methylpropane sulfonic acid) and the pH and contact angle value were verified and compared to commercial ScotchBond primer. Adhesive resin with HEAA and TMA (TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%) showed lower conversion and polymerization rate after 40 s of light activation. Higher conversion (up to 60%) was found for BisGMA/HEAA/TMA and BisGMA/HEMA adhesive resin without significant difference between adhesive resin, p>0.05. UTS (BisGMA/HEMA- 67.7 ±5 MPa e BisGMA/HEAA/TMA- 60.5 ±7 MPa), immediate μTBS (BisGMA/HEMA- 57 ± 14 MPa e BisGMA/HEAA/TMA- 53.1 ±15 MPa), ΔKHN (BisGMA/HEMA- 56 ± 7 e BisGMA/HEAA/TMA- 64 ±4) and contact angle (BisGMA/HEMA- 39.5 ±9 e BisGMA/HEAA/TMA- 46.7 ±15) showed no statistical difference, p>0.05. The experimental primer presented more acidity pH (2.7) and lower contact angle (18.5 ±5) when compared to commercial primer (pH- 4 e θ- 33.5 ±4). A new acrylamide based-primer was formulated and the presence of the new tris(methacrylamide) monomer (TMA) was enable the preparation of a 3-step etch-and-rise adhesive system without HEMA monomer.

O objetivo deste estudo é caracterizar a influência da relação dos componentes hidrófilos de uma resina adesiva experimental em suas propriedades intrínsecas. Para isso, foram adicionados a um sistema polimérico (BisGMA, BisEMA e TEGDMA), diferentes concentrações de HEMA totalizando quatro grupos: 0%, 15%, 30% e 50% de HEMA. As misturas foram caracterizadas quanto à cinética de polimerização através de Fourier Transformed Infrared (FTIR) em tempo real e a taxa de polimerização (RP) foi calculada através da análise de regressão não-linear; resistência coesiva à tração utilizando-se uma máquina de ensaios mecânicos com corpos de prova em forma de ampulheta com área de constrição de 1mm2; e, sorção e solubilidade baseada na normatização ISO 4049. Houve correlação entre o tempo de fotoativação e o grau de conversão para todos os grupos (r2>0.98). O grupo 0% apresentou RPMÁX elevada nos segundos iniciais de fotoativação com, no entanto, uma brusca redução. As adições de 30% e 50% de HEMA determinaram uma RPMÁX do copolímero menor que em 15% e 0%. Não houve diferença entre o grau de conversão final (60s) entre os grupos de 0, 15 e 30%, sendo 59,7%, 61,4% e 55,2% respectivamente. As adições de 30% e 50% de HEMA determinaram a redução da resistência coesiva dos polímeros (p<0,05) quando comparado aos grupos 0% e 15%. Grupos com maiores adições de HEMA apresentaram maiores graus de sorção e solubilidade (p<0,05). Maiores adições de HEMA resultaram em polímeros mais propensos à degradação. / The aim of this study was to characterize the influence of hydrophilic monomers ratios on an experimental adhesive resin properties. Four groups, with 0, 15, 30 and 50% (wt) addiction of HEMA were prepared. Comonomer blends (BisGMA, BisEMA, TEGDMA and HEMA) were analyzed with kinetics of polimerization real time FTIR and the rate of polymerization was calculated by non-linear regression analysis. Ultimate tensile strength was tested using a universal testing machine (Emic) with hour-glass shaped specimens with cross-sectional area 1mm2. Water sorption and solubility tests were based on ISO 4049. For All groups was a positive and strong correlation between light-activation time and degree of conversion (r2>0,98). Group 0% of HEMA showed higher RPMÁX in early seconds with a suddenly decrease. Additions of 30% and 50% of HEMA determined a RPMÁX lower than the comonomer blends with 15% and 0% of HEMA. There was no statistical difference of final degree of conversion of the tested groups, varying from 63% to 67% . The additions of 30% and 50% of HEMA determined reduction of ultimate tensile strength of the polymers formed (p<0,05) when compared to 0% and 15 % of HEMA. Groups with higher additions of HEMA showed higher water sorption and solubility (p<0,05). Higher additions of HEMA resulted in polymers more prone to degradation.

O objetivo deste estudo foi sintetizar e caracterizar monômeros metacrilamidas, desenvolver, caracterizar e avaliar as propriedades de sistemas adesivo convencional de três passos. Foram sintetizadas bis(metacrilamida)s e tris(metacrilamida) e caracterizadas por espectroscopia de Infravermelho por Transformada de Fourier (FTIR), Ressonância Magnética Nuclear (RMN) de 1H e 13C, por Cromatografia Líquida de Alta Eficiência com Espectrometria de Massas (UHPLC-QTOF-MS) e Calorimetria Exploratória Diferencial Modulada (MDSC). Quatro bis(metacrilamidas), (1) N,N’-(propane-1,3-diyl)bis(N-ethyl-2-methylacrylamide), (2) N,N’-(octane-1,8-diyl)bis(2-methylacrylamide), (3) N,N’-(butane-1,4-diyl)bis(2-methacrylamide) e (4) N,N’-(1,4 phenylene)bis(2-methylacrylamide)monômero (1), (2), (3) e (4), e uma tris(metacrilamida) TMA, foram sintetizadas. Pela análise de FTIR-ATR foram observadas as bandas correspondentes ao estiramento do grupo C=O (1660 cm-1), C=C (1610 cm-1), N-H (3300 cm-1) e C-N (1520 cm-1). As análises de RMN identificaram a presença das ligações duplas referentes aos grupos metacrilamidas em deslocamentos químicos entre 5,3 e 5,8 ppm para 1H e entre 120 e 140 ppm para 13C.Os valores de massa exata m/z foram: 267,2068, 281,2222, 225,1595, 245,1283 e 351,2385 g/mol para os monômeros (1), (2), (3), (4) e TMA, respectivamente. A cinética de polimerização do TMA e dos adesivos experimentais contendo 2-hidroxietil acrilamida (HEAA) ou 2-hidroxietil metacrilato (HEMA) com as seguintes formulações foram investigadas por meio de DSC-PCA, n=3: TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%, TMA50%/HEMA50%, BisGMA/HEAA/TMA e BisGMA/HEMA.Características e propriedades mecânicas das resinas adesivas BisGMA/HEAA/TMA e BisGMA/HEMA foram avaliadas por resistência coesiva (UTS, n=5), degradação em solvente (ΔKHN, n=5), ângulo de contato (n=5), microtração (μTBS, n=20) e análise de fratura. Um primer a base de acrilamidas foi desenvolvido (H2O/HEAA/AMPS) (2-acrylamida-2-methilpropano ácido sulfônico) para ser utilizado no grupo experimental com metacrilamidas. Os valores de pH e ângulo de contato do primer experimental foram comparados com o primer do ScotchBond Multi-purpose (grupo controle). O monômero (1) resultou em um monômero amarelo claro de baixa viscosidade, entretanto, não apresentou foto ou termopolimerização. A energia de ativação determinada pelo método de Kissinger foi – 165,8 kJmol-1; -182,7 kJmol-1 e -156,7 kJmol-1 para os monômeros (2), (3) e (4), respectivamente. Sistemas adesivos convencionais de três passos a base de metacrilamidas e a base de metacrilatos foram desenvolvidos. Resinas adesivas contendo somente HEAA e TMA (TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%) apresentaram grau de conversão abaixo de 40% após 40 s de fotoativação. Alto grau de conversão (acima de 60%) só foi encontrado para as resinas adesivas BisGMA/HEAA/TMA e BisGMA/HEMA e sem diferença significativa entre elas, p>0,05. Os valores de UTS (BisGMA/HEMA- 67,7 ±5 MPa e BisGMA/HEAA/TMA- 60,5 ±7 MPa), μTBS (BisGMA/HEMA- 57 ± 14 MPa e BisGMA/HEAA/TMA- 53,1 ±15 MPa) e ângulo de contato (BisGMA/HEMA- 39,5 ±9 e BisGMA/HEAA/TMA- 46,7 ±15) não apresentaram diferença estatística, p>0.05. O primer experimental apresentou um valor pH mais baixo (2,7) bem como de ângulo de contato (18,5 ±5) em relação ao comercial (pH-4 e θ-33,5 ±4). A síntese proposta para os monômeros (1), (2), (3), (4) e TMA foi caracterizada nesse trabalho. Um primer somente com acrilamidas foi desenvolvido e a presença do novo monômero TMA na resina adesiva BisGMA/HEAA permitiu a formulação de um sistema adesivo convencional de três passos sem a presença do monômero HEMA. / The aim of this study was synthesized and characterizes methacrylamides monomers, development, characterizer and evaluated the properties of 3-step etch-and-rise adhesive system. Bis(methacrylamide)s and tris(methacrylamide) were synthesized. The monomer structures were confirmed by 1H and 13C Nuclear Magnetic Resonance (NMR), Fourier Transform Infrared Spectroscopy (FTIR-ATR), Ultra-high liquid chromatography quadrupole time of flight mass spectrometry (UHPLC-QTOF-MS) and modulated differential scanning calorimetry (mDSC). Four bis(methacrylamide)s monomers (1) N,N’-(propane-1,3-diyl)bis(N-ethyl-2-methylacrylamide), (2) N,N’-(octane-1,8-diyl)bis(2-methylacrylamide), (3) N,N’-(butane-1,4-diyl)bis(2-methacrylamide) and (4) N,N’-(1,4 phenylene)bis(2-methylacrylamide) and one tris(methacrylamide), TMA, were synthesized. All IR spectra of the monomers showed the C=C axial deformation at 1610 cm-1. The 1H NMR spectra the olefinic hydrogens were observed at 5.3 an 5.8 ppm and in the 13C NMR, the vinylic carbons at 120 and 140 ppm. The exact m/z values were: 267.2068, 281.2222, 225.1595, 245.1283 and 351.2385 g/mol for monomers (1), (2), (3), (4) and TMA respectively. Monomer (1) not presented photo (DSC-PCA) or thermal polymerization. The activation energy determined using Kissinger methodology was: - 165.8 kJmol-1; -182.7 kJmol-1 and -156.7 kJmol-1 for monomers (2), (3) and (4), respectively. 3-step adhesive systems with methacrylamides and methcrylates were development. Kinetics of photopolymerization of TMA and experimental adhesive resin containing 2-hydroxyethylacrylamide (HEAA) or 2-hydroxyethylmethacrylate (HEMA) in the following formulations: (TMA 33%/HEAA 66%, TMA 50%/HEAA 50%, TMA66%/HEAA33%, TMA50%/HEMA50%, BisGMA66%/HEAA24%/TMA10% and BisGMA66%/HEMA33%) were investigated through DSC-PCA. Characteristics and mechanical properties for BisGMA 66%/HEAA 24%/TMA 10% and BisGMA 66%/HEMA 33% adhesives were evaluated with ultimate tensile strength (UTS, n=5), softening in solvent (ΔKHN, n=5), contact angle (n=5), microtensile bond strength (μTBS, n=20) and failure analysis. A primer was also formulated with H2O/HEAA/AMPS (2-acrylamido-2-methylpropane sulfonic acid) and the pH and contact angle value were verified and compared to commercial ScotchBond primer. Adhesive resin with HEAA and TMA (TMA33%/HEAA66%, TMA50%/HEAA50%, TMA66%/HEAA33%) showed lower conversion and polymerization rate after 40 s of light activation. Higher conversion (up to 60%) was found for BisGMA/HEAA/TMA and BisGMA/HEMA adhesive resin without significant difference between adhesive resin, p>0.05. UTS (BisGMA/HEMA- 67.7 ±5 MPa e BisGMA/HEAA/TMA- 60.5 ±7 MPa), immediate μTBS (BisGMA/HEMA- 57 ± 14 MPa e BisGMA/HEAA/TMA- 53.1 ±15 MPa), ΔKHN (BisGMA/HEMA- 56 ± 7 e BisGMA/HEAA/TMA- 64 ±4) and contact angle (BisGMA/HEMA- 39.5 ±9 e BisGMA/HEAA/TMA- 46.7 ±15) showed no statistical difference, p>0.05. The experimental primer presented more acidity pH (2.7) and lower contact angle (18.5 ±5) when compared to commercial primer (pH- 4 e θ- 33.5 ±4). A new acrylamide based-primer was formulated and the presence of the new tris(methacrylamide) monomer (TMA) was enable the preparation of a 3-step etch-and-rise adhesive system without HEMA monomer.

O objetivo deste estudo é caracterizar a influência da relação dos componentes hidrófilos de uma resina adesiva experimental em suas propriedades intrínsecas. Para isso, foram adicionados a um sistema polimérico (BisGMA, BisEMA e TEGDMA), diferentes concentrações de HEMA totalizando quatro grupos: 0%, 15%, 30% e 50% de HEMA. As misturas foram caracterizadas quanto à cinética de polimerização através de Fourier Transformed Infrared (FTIR) em tempo real e a taxa de polimerização (RP) foi calculada através da análise de regressão não-linear; resistência coesiva à tração utilizando-se uma máquina de ensaios mecânicos com corpos de prova em forma de ampulheta com área de constrição de 1mm2; e, sorção e solubilidade baseada na normatização ISO 4049. Houve correlação entre o tempo de fotoativação e o grau de conversão para todos os grupos (r2>0.98). O grupo 0% apresentou RPMÁX elevada nos segundos iniciais de fotoativação com, no entanto, uma brusca redução. As adições de 30% e 50% de HEMA determinaram uma RPMÁX do copolímero menor que em 15% e 0%. Não houve diferença entre o grau de conversão final (60s) entre os grupos de 0, 15 e 30%, sendo 59,7%, 61,4% e 55,2% respectivamente. As adições de 30% e 50% de HEMA determinaram a redução da resistência coesiva dos polímeros (p<0,05) quando comparado aos grupos 0% e 15%. Grupos com maiores adições de HEMA apresentaram maiores graus de sorção e solubilidade (p<0,05). Maiores adições de HEMA resultaram em polímeros mais propensos à degradação. / The aim of this study was to characterize the influence of hydrophilic monomers ratios on an experimental adhesive resin properties. Four groups, with 0, 15, 30 and 50% (wt) addiction of HEMA were prepared. Comonomer blends (BisGMA, BisEMA, TEGDMA and HEMA) were analyzed with kinetics of polimerization real time FTIR and the rate of polymerization was calculated by non-linear regression analysis. Ultimate tensile strength was tested using a universal testing machine (Emic) with hour-glass shaped specimens with cross-sectional area 1mm2. Water sorption and solubility tests were based on ISO 4049. For All groups was a positive and strong correlation between light-activation time and degree of conversion (r2>0,98). Group 0% of HEMA showed higher RPMÁX in early seconds with a suddenly decrease. Additions of 30% and 50% of HEMA determined a RPMÁX lower than the comonomer blends with 15% and 0% of HEMA. There was no statistical difference of final degree of conversion of the tested groups, varying from 63% to 67% . The additions of 30% and 50% of HEMA determined reduction of ultimate tensile strength of the polymers formed (p<0,05) when compared to 0% and 15 % of HEMA. Groups with higher additions of HEMA showed higher water sorption and solubility (p<0,05). Higher additions of HEMA resulted in polymers more prone to degradation.

ABSTRACT Covalent Immobilization of Glucose Isomerase on Poly (2-hydroxyethyl methacrylate) Particles Yildiz, Hakan &Uuml / mit M.S., Department of Chemistry Supervisor: Prof. Dr. Nesrin Hasirci July 2004, 54 pages In this study, poly (2-hydroxyethyl methacrylate), P(HEMA), particles were prepared by suspension polymerization of the monomer 2-hydroxyethyl methacrylate with addition of ethylene glycol dimethyacrylate, EGDMA, as cross linker. Glucose isomerase, GI, enzyme was covalently immobilized on the prepared P(HEMA) particles after activation of the particles with cyanuric chloride. The activities of the free and immobilized enzymes were measured with Ethanol-Carbazole method. The immobilization of GI on P(HEMA) particles promoted enzyme stability and as a result, the enzyme became more stable to temperature, storage, and reuse. For maximum substrate conversion, optimum temperature was determined as 70 oC for free GI and this value shifted to 60 oC for immobilized enzyme. Optimum pH for maximum substrate conversion was found to be 7.0 for free GI and 8.0 for immobilized GI. The change of enzyme activity with substrate concentration were determined to calculate Km and Vmax values of the free and immobilized enzymes. Km values were found to be 1.7x10-2 mol/L and 3.1x10-1 mol/L while Vmax values were 1.01x10-4 mol/L.min, 1.65x10-3 mol/L.min for free and immobilized GI, respectively. Reuse capability of immobilized GI on P(HEMA) particles was measured and compared with commercial GI. Both systems retained 80 % of their original activities after 40th use, within 6 days. The change of enzyme activities upon storage were detected at certain time intervals for the samples stored in buffer solution at 4 oC. Immobilized enzyme was retained 60% of its original activitiy in 60 days of storage at 4 oC. Immobilized GI and commercial GI both retained 90% of their activities under continuous flow after 180 mL of substrate solution passed through the column.

Due to sensor -tissue interactions, currently none of the commercially available glucose sensors are capable of continuous, reliable monitoring of glucose levels during long-term implantation. In order to improve the lifetime of implanted glucose sensors, two series of biocompatible novel hydrogel coatings were designed, synthesized and the physical properties were measured. Different hydrogels with various 2,3-dihydroxypropyl methacrylate (DHPMA) compositions were coated onto glucose sensors. Results show that the higher freezable water content, swelling rate and uniform porosity that resulted from high DHPMA content increased the sensitivity and shortened the response time of glucose sensors. The linear range of a glucose sensor coated only with hydrogel is short, however, the range can be improved by coating the epoxy- polyurethane (PU) with a layer of hydrogel. Since the hydrogel minimizes the fibrosis and inflammation, it shows promise for use in implantable glucose sensors. However, the in vivo experiment shows only 25% of sensors still worked after 4 weeks. In order to overcome problems present in the first series of experiments, another series of novel hydrogels with various N-vinyl pyrolidone (VP) contents was developed. This study has provided a feasible approach to design and select the properties of the copolymer for coating implantable biosensors. The in vivo experiments demonstrate that a hydrogel coating significantly improved the performance of implanted glucose sensors. In order to suppress the acute inflammation caused by the surgery, dexamethasone-21 phosphate disodium salt (DX-21) was incorporated to a series of poly (HEMA-DHPMA-VP) hydrogels to investigate the drug delivery in vitro. All hydrogels showed a high initial release, followed by slow, long term release during the next hours to days. This release pattern is believed to be optimum for implanted glucose sensors suppressing the acute and chronic inflammation. Water structures in hydrogels swollen in different media water, PBS and DX-21 solution were also investigated. 1HEMA:1DHPMA copolymer and VP-HEMA-DHPMA copolymers imbibed higher freezable water fractions in DX-21 solution. The ratio of transporting water mass to DX-21 mass is 9.6 which is independent of the hydrogel composition.

<p>In the presented work, immunosensors for detection of Aflatoxin B1 based on different immobilization platforms were studied. Synthesis of an electroactive hydrogel was also carried out. Aflatoxins are a group of mycotoxins that have deleterious effects on humans and are produced during fungal infection of plants or plant products. Electrochemical immunosensor for the determination of Aflatoxin B1 (AFB1) was developed with anti-aflatoxin B1 antibody immobilized on Pt electrodes modified with polyaniline (PANi) and polystyrene sulphonic acid (PSSA). Impedimetric analysis shows that the electron transfer resistances of Pt/PANi-PSSA electrode, Pt/PANi-PSSA/AFB1-Ab immunosensor and Pt/PANi-PSSA/AFB1-Ab incubated in BSA were 0.458, 720 and 1066 k&Omega / , respectively. These results indicate that electrochemical impedance spectroscopy (EIS) is a suitable method for monitoring the change in electron-transfer resistance associated with the immobilization of the antibody. Modelling of EIS data gave equivalent circuits which showed that the electron transfer resistance increased from 0.458 k&Omega / for Pt/PANi-PSSA electrode to 1066 k&Omega / for Pt/PANi-PSSA/AFB1-Ab immunosensor, indicating that immobilization of the antibody and incubation in BSA introduced an electron transfer barrier. The AFB1 immunosensor had a detection limit of 0.1 mg/L and a sensitivity of 869.6 k &Omega / L/mg.</p>

A strategic research area today is development of polymericproducts made from renewable sources. The ways of utilizingrenewable sources studied in this thesis are using 1)prepolymers obtained by steam treatment of wood and 2) monomersobtainable by fermentation of agricultural products. Novel hemicellulose-based hydrogels were prepared by usingprepolymers obtained from steam treatment of spruce.Hemicellulose was first modified with well-defined amounts ofmethacrylic functions. Hydrogels were then prepared by radicalpolymerization with 2-hydroxyethyl methacrylate orpoly(ethylene glycol) dimethacrylate to form hydrogels. Theradical polymerization reaction was carried out in water usinga redox initiator system. The hydrogels were in generalelastic, soft and easily swollen in water. Frequency sweeptests indicated that the hydrogel system displayed prevailingsolid-like behavior. Comparison of the hemicellulose-basedhydrogels with pure poly(2-hydroxyethyl methacrylate)-basedhydrogels showed that it was possible to preparehemicellulose-based hydrogels with properties similar to thoseof pure poly(2-hydroxyethyl methacrylate)-based hydrogels. Polyester-based materials were prepared by using themonomers 1,3- propanediol and succinic acid obtainable byfermentation. α,ω-Dihydroxyterminatedoligomeric polyesters produced by the thermal polycondensationof 1,3-propanediol and succinic acid were chain-extended toobtain sufficiently high molecular weight. Depending on thechain-extension technology adopted, poly(ester carbonate)s orpoly(ester urethane)s were obtained. In the case of poly(estercarbonate)s, the chain-extended products ofα,ω-dihydroxyterminated oligomeric copolyesters werealso produced using 1,3-propanediol/1,4-cyclohexanedimethanol/succinic acid mixtures toimprove thermal and mechanical properties. Segmented poly(esterether carbonate)s fromα,ω-dihydroxyterminated oligo(propylenesuccinate)s and poly(ethylene glycol) were also synthesized toincrease the hydrophilicity. Molecular weights and polydispersity were analyzed by SECfor all materials. Their structures were also identified by NMRspectroscopy (1H NMR and 13C NMR). All characterizations werein agreement with the proposed structures. Thermal parameterswere characterized by DSC. Tensile testing anddynamic-mechanical tests were performed and in additionpreliminary processing trials were carried out in some cases.The results demonstrate the feasibility of using monomersderived from renewable sources to build up new polymericstructures endowed with a variety of physical and mechanicalproperties.