Orientador: Idomeo Bonetti Filho / Banca: Renato de Toledo Leonardo / Banca: Lizeti Toledo de Oliveira Ramalho / Banca: Celso Kenji Nishiyama / Banca: Ivaldo Gomes de Moraes / Resumo: Seguindo critérios da ISO/FDI e ADA/ANSI, avaliou-se a resposta tecidual ao cimento EndoREZ®, um cimento experimental derivado do polímero de mamona (Poliquil®), Endofill® e Sealapex®. Tubos de polietileno contendo os cimentos e o grupo controle, foram implantados em duas lojas cirúrgicas preparadas no tecido subcutâneo dorsal de 26 camundongos Mus muscullus albinus machos. Após 7 e 50 dias os animais foram mortos e removidos cirurgicamente os fragmentos teciduais, os quais foram fixados por 96 horas em solução de formol tamponado e processado através das técnicas histológicas de rotina. Seções de 6mm de espessura foram coradas com Hematoxilina e Eosina e Tricrômico de Mallory, e avaliadas com o emprego de microscópio óptico comum (Carl Zeiss - Jenaval). As análises mostraram, discreto ou moderado infiltrado inflamatório dos materiais testados durante os dois períodos, com exceção do cimento experimental derivado do polímero de mamona, o qual apresentou ótimo comportamento biológico mostrando seu potencial para o desenvolvimento de um novo material obturador de canais radiculares. / Abstract: According to ISO/FDI and ADA/ANSI criteria, it was evaluated tissue responses of EndoREZ, Castor Oil Polymer Sealer, Endofill and Sealapex. Polyethylene tubes containing the cements and control group, were implanted into two surgical pockets prepared in the dorsal subcutaneous tissue of 26 rats. After 7 and 50 days, the implants were surgically removed, fixed for 96 hours in buffered formalin and processed using routine histological techniques. Sections of 6 mm were stained with Hematoxylin and Eosin and evaluated under light microscopy. Analysis revealed mild and moderate inflammatory infiltrate for all materials in both periods, exception to Castor Oil Polymer Sealer that presented better biological behavior. / Doutor

Orientador: Elcio Marcantonio Junior / Banca: Lizeti Toledo de Oliveira Ramalho / Banca: Luís Antônio Violin Dias Pereira / Resumo: O objetivo desta pesquisa foi desenvolver biomateriais à base de quitosana e de cloridrato de quitosana (géis), e avaliar radiográfica e histológicamente seu uso na correção de defeitos ósseos críticos na calvária de ratos. Para tal, defeitos de 8mm de diâmetro foram criados cirurgicamente na calvária de 50 ratos Holtzman (Rattus norvegicus) e preenchidos com coágulo sanguíneo (controle), quitosana com baixo peso molecular, quitosana com alto peso molecular, cloridrato de quitosana com baixo peso molecular e cloridrato de quitosana com alto peso molecular, totalizando 10 animais, divididos em dois períodos experimentais (15 e 60 dias), para cada biomaterial. Na avaliação radiográfica foram utilizadas duas radiografias digitais dos crânios dos animais, feitas logo após a criação do defeito ósseo e no momento do sacrifício, que forneceram a densidade óssea radiográfica na área do defeito. A avaliação histológica pautou-se na descrição da morfologia dos tecidos neoformados na região do defeito. Os resultados radiográficos e histológicos indicaram que, em todos os grupos, a neoformação óssea ocorreu apenas nas bordas do defeito, e que, exceto no grupo controle, graus variados de inflamação foram observados. Conclui-se que, dentro das condições experimentais deste estudo, os biomateriais de quitosana e de cloridrato de quitosana não foram capazes de promover neoformação óssea nos defeitos ósseos críticos criados em calvária de ratos. / Abstract: The purpose of this study was to develop chitosan and chitosan chloride biomaterials (gels), and to make a histological and radiographic evaluation of their use in the correction of critical size bone defects made in rat's calvaria. Bone defects of 8mm in diameter were surgically created in the calvaria of 50 Holtzman (Rattus norvegicus) rats and filled with blood clot (control), low molecular weight chitosan, high molecular weight chitosan, low molecular weight chitosan chloride and high molecular weight chitosan chloride, numbering 10 animals, divide into two experimental periods (15 and 60 days), for each biomaterial. For the radiographic evaluation, it were made two digital radiographs of animal's skulls, right after the creation of the bone defect and at the moment of the sacrifice, providing the radiographic bone density in defect's area. Histological evaluation was made based on the morphology of the new formed tissues in defect's region. Radiographic and histological results indicated that, in all groups, new bone formation occurred only near to the defect's border, and, except in control groups, assorted degrees of inflammation could be seen. We conclude that chitosan and chitosan chloride biomaterials were not able to promote new bone formation incritical size defects made in rat's calvaria. / Mestre

Orientador: Elisa Maria Aparecida Giro / Banca: Carlos Alberto de Souza Costa / Banca: João Eduardo Gomes Filho / Resumo: Para que um material seja indicado como retrobturador, é desejável que apresente várias propriedades, entre elas, intensa atividade antibacteriana e aceitável biocompatibilidade. O presente trabalho teve como objetivo avaliar, in vitro, o efeito antibacteriano e citotóxico do Agregado de Trióxido Mineral (MTA-Branco, Angelus® Indústria de Produtos Odontológicos Ltda., Londrina, PR, Brasil), associado à solução de clorexidina (CHX) em diferentes concentrações. Para a análise da atividade antibacteriana foi empregado o método de difusão em ágar Müller-Hinton e foram utilizados os microrganismos Staphylococcus aureus (ATCC - 25923), Pseudomonas aeruginosa (ATCC - 27853), Escherichia coli (ATCC - 25922) e Enterococcus faecalis (ATCC - 29262). Em placas de Petri contendo a suspensão bacteriana com meio de cultura, foram preparados poços com 4 mm de diâmetro e 5 mm de profundidade, os quais foram preenchidos com o cimento MTA preparado com água destilada estéril (G1-controle) ou com solução de CHX nas concentrações de 0,06% (G2), 0,12% (G3), 0,2% (G4) e 1% (G5). As placas foram incubadas em condições de aerobiose a 37ºC por 24 horas e os halos de inibição do crescimento bacteriano foram medidos em milímetros. Para avaliar a citotoxicidade do MTA associado às diferentes concentrações de CHX, o metabolismo e a morfologia celular foram avaliados por meio do teste do MTT e da análise da microscopia eletrônica de varredura, respectivamente. Corpos de prova de 4 mm de diâmetro e 2 mm de espessura foram preparados com as associações de materiais e imersos em 1,1 mL de meio de cultura (DMEM) por 24 horas ou por 7 dias para obtenção dos extratos. DMEM completo (grupo controle - G6) e os extratos de cada grupo foram aplicados sobre cultura de fibroblastos do ligamento periodontal de camundongos, os quais foram incubados... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: An ideal root-end filling material should present several properties, including antibacterial activity and biocompatibility. The aim of this in vitro study was to investigate the antibacterial and cytotoxic effects of Mineral Trioxide Aggregate (MTA-White, Angelus® Indústria de Produtos Odontológicos Ltda., Londrina, PR, Brasil) mixed with different concentrations of chlorhexidine (CHX). For antibacterial activity analysis diffusion method on Müller-Hinton agar was employed. The bacterial strains used were Staphylococcus aureus (ATCC - 25923), Pseudomonas aeruginosa (ATCC - 27853), Escherichia coli (ATCC - 25922) e Enterococcus faecalis (ATCC - 29262). In Petri plates containing culture medium with bacterial suspension, wells were made measuring 4 mm in diameter and 5 mm in depth. The wells were completely filled with MTA mixed with sterile distilled water (G1) or with different concentrations of CHX: 0.06 % (G2), 0.12% (G3), 0.2% (G4) and 1% (G5). The plates were incubated in aerobic conditions at 37ºC for 24 hours, and then the diameters of the zones of bacterial inhibition were measured. To evaluate the cytotoxicity of the MTA mixed with different concentrations of CHX, the cell metabolism and cells morphology were evaluated by the MTT assay and scanning electron microscopy, respectively. Round-shaped samples measuring 4 mm in diameter and 2 mm in thickness were prepared with the association of materials and immersed in 1.1 mL of culture medium (DMEM) and incubated for 24 hours or 7 days. The extracts from every sample and pure DMEM (control group - G6) were applied on mouse periodontal ligament fibroblast cells culture and incubated at 37ºC with 5% CO2 and 95% air for 24 hours. The addition of CHX 1% to MTA increased the antibacterial activity of the cement against S. aureus (p <0.05). There was no statistically significant difference... (Complete abstract click electronic access below) / Mestre

Os objetivos do trabalho foram caracterizar as biocerâmicas a base de hidroxiapatita (HA), -fosfato tricálcio (TCP) e o compósito HA/TCP na proporção 60:40 pela análise microestrutural e por Espectrometria de Energia Dispersiva de Raios-X (EDS) ao microscópio eletrônico de varredura, e comparar histomorfometricamente o processo de reparo ósseo de defeitos cranianos perenes tratados com essas três diferentes cerâmicas e o enxerto autógeno nos períodos de 0, 30, 90 e 180 dias pós-operatório. Microestruturalmente, a HA apresentou superfície irregular com concavidades, fendas e grande quantidade de microporos; o TCP exibiu superfície irregular e baixa resistência mecânica; a HA/TCP apresentou superfície lisa com raras concavidades e ausência de porosidades. As cerâmicas estavam constituídas pelos elementos O, P e Ca, sendo que a relação Ca/P seguiu a ordem: HA/TCP > HA > TCP. Morfometricamente, o volume inicial enxertado foi aproximadamente de 60 mm3 para todos os tratamentos. Entre 30 e 180 dias, o volume total da região do enxerto (Vtre) e do material enxertado (VM) manteve-se constante nos defeitos tratados com osso autógeno (Vtre = 53,61 mm³, VM =20,11 mm³), HA (Vtre = 60,13mm³, VM = 32,72 mm³), HA/TCP (Vtre = 57,60mm³, VM = 31,61 mm³), TCP (Vtre = 38,27mm3, VM = 10,07 mm³). Aos 30 dias, o volume de tecido ósseo no grupo autógeno foi de 22,06 mm³, ocupando quase a totalidade dos espaços entre as partículas, enquanto que, nos grupos HA e HA/TCP foi de apenas 7,62 mm³, sendo parte dos espaços (19,10 mm3), constituído por tecido conjuntivo fibroso. Entre 30 e 180 dias, o volume de tecido ósseo não apresentou diferenças estatísticas significantes (p>0,05). O número absoluto de osteoblastos e osteócitos no grupo autógeno foi, respectivamente, de 118,57 x102 e 178,57 x102 células, aos 30 dias, diminuindo 40,16% e 29,09% aos 90 dias, decorrente do processo de remodelação óssea. Nos grupos HA e HA/TCP, o número absoluto de osteoblastos e osteócitos foram similares e permaneceram constantes dos 30 aos 180 dias, com uma média de 39,13 x102 e 132,20 x102 células, respectivamente. Com relação ao grupo TCP, o volume de tecido ósseo foi de 4,16mm3 aos 30 dias e aumentou 70,27% aos 90 dias, passando a ocupar 13,93 mm³ do Vtre. Concomitantemente, o número absoluto de osteoblastos e osteócitos passaram, respectivamente, de 5,64 x102 e 37,01 x102 células, aos 30 dias, para 67,46 x102 e 166,13 x102 células, aos 90 dias. Baseados nos resultados aqui obtidos, concluímos que, no modelo experimental de defeitos cranianos em ratos, o enxerto autógeno é ainda, o padrão ouro para reconstrução óssea, enquanto que, dentre as cerâmicas analisadas, o TCP é o melhor biomaterial ósseo-substituto. / The aim of the work was to characterize the ceramics: hydroxyapatite (HA), -tricalcium phosphate (TCP) and the composite HA/TCP (60:40) through scaning electron microscope - energy dispersive system (SEM-EDS) analysis and to compare histomorphometrically the repair of perennial skull defects treated with these three different ceramics and autogenous graft at 0, 30, 90 and 180 days post-operatively. Microstructurally, HA showed irregular surface with concavities, cracks and several pores. TCP also presented irregular surface and low mechanical strength. HA/TCP showed smooth surface with rare concavities and pores. The ceramics were constituted by the elements O, P and Ca, and the relation Ca/P follow the order: HA/TCP > HA > TCP. Morphometrically, the initial graft volume was approximately 60 mm3 for all treatments. Between 30 and 180 days, the total volume of the grafted area (Vtga) and of the implanted material (Vim) were constant in the following groups: autogenous (Vtga = 53,61 mm³; Vim = 20,11 mm3), HA (Vtga = 60,13mm³; Vim = 32,72 mm3), HA/TCP (Vtga = 57,60mm³; Vim = 31,61 mm3) and TCP (Vtga = 38,27mm³; Vim = 10,07 mm3). At 30 days, the bone tissue volume was of 22,06 mm3 in the autogenous group, occupying totally the space among the particles, while in the HA and HA/TCP groups was only 7,62 mm3. At 30 and 180 days, the bone tissue volume did not show statistical differences (p>0.05). The total number of osteoblasts and osteocytes in the autogenous group was respectively, 118, 57 x 102 and 178,57 x 102 cells, at 30 days, reducing 40,16% e 29,09%, at 90 days, due to bone remodeling. In the HA and HA/TCP groups, a similar and stable number of osteoblasts and osteocytes were observed during all periods, with mean of 39,13 x 102 and 132,20 x 102 cells, respectively. In the TCP group, the bony tissue volume was of 4,16 mm3, at 30 days, increasing 70,27% at 90 days. Concomitantly, the total number of osteoblasts and osteocytes was respectively, 5,64 x 102 and 37,01 x 102 cells, at 30 days, achieving 67,46 x 102 and 166,13 x 102 cells, at 90 days. Based in the results obtained here, we concluded that in the rat skull defect model the autogenous bone is yet the good standard graft material for bone reconstruction and that among the tested ceramics, the TCP is the best bone graft substitute.

Com a evolução do Biomateriais houve melhorias nas opções de tratamentos e atualmente são utilizados em substituição de partes do corpo que foram perdidas e promovem a recuperação de funções biológicas. Dentre eles existem as chamadas Biocerâmicas, que incluem alumina, zircônia e derivadas de fosfato de cálcio. A hidroxiapatita possui composição e estrutura minerais semelhantes à fase mineral óssea e apresenta como propriedades a biocompatibilidade, osteocondutividade e bioatividade. O trabalho avaliou a viabilidade celular em cerâmica de hidroxiapatita experimental de origem bovina em comparação com dois tipos de zircônia comerciais e liga de titânio comercialmente puro, para que futuramente possa ser utilizada como material base para implantes dentários. A avaliação in vitro foi realizada por meio de testes nos quais células pré-osteoblásticas cultivadas de linhagem murina MC3T3-E1 foram colocadas em contato indireto e direto com estes materiais. Para viabilidade celular (n=8) foram feitos testes de ensaio MTT e Cristal Violeta em duplicata e após 24, 48 e 72 horas os níveis de absorbâncias foram analisados por meio de espectrofotometria no leitor de Elisa. Para as analises por microscopia eletrônica de varredura (n=6) as células foram plaqueadas diretamente sobre as superfícies dos discos, fixadas em vapor de tetróxido de ósmio 2% após 24 e 48 horas, seguido da metalização após 48 horas da fixação das células para análise em Microscópio Eletrônico de Varredura. Os resultados para viabilidade indireta foram submetidos ao teste paramétrico ANOVA, seguido de teste de Tukey (p<0,05). Tanto no teste de MTT quanto no Cristal Violeta, de acordo com o grupo controle positivo, todos os grupos apresentaram resultados satisfatórios. A cerâmica de hidroxiapatita não apresentou diferença estatística significante, demonstrando não ser um material citotóxico. Pelas imagens geradas no MEV do teste de viabilidade direta, verificou-se que houve adesão e proliferação das células nos dois períodos sobre as superfícies dos materiais. Portanto, pode-se afirmar que a cerâmica de hidroxiapatita apresentou-se como um material biocompatível. / With the evolution biomaterials there were improvements in treatment options, and are currently used in replacement body parts that were lost and promote the recovery of biological functions. Among them are the bioceramic which include alumina, zirconia and calcium phosphate derivative. Hydroxyapatite has mineral composition and structure similar to bone mineral phase and can be used as a biomaterial having biocompatibility, osteoconductivity and bioactivity. The study evaluated the cell viability in experimental hydroxyapatite ceramic bovine compared the two types of commercial zirconia and titanium alloy commercially pure, so that in future it can be used as base material for dental implants. In vitro evaluation was carried by means of tests in which the pre-osteoblastic cells MC3T3-E1 murine lineage cultured were placed in indirect and direct contact with these materials. For cell viability (n=8) were carried MTT assay and Crystal Violet tests in duplicate and after 24, 48 and 72 hours the absorbance levels were analyzed by spectrophotometry Elisa reader. For analysis by Scanning Electron Microscope variable pressure (n = 6) cells were plated directly on the discs surfaces, fixed in osmium tetroxide steam 2% after 24 and 48 hours, followed by metallization after 48 of cells fixation. The results for the cell viability were submitted to parametric test ANOVA, followed by Tukey test (p<0.05). Both in the MTT assay as Crystal Violet all groups exhibited satisfactory results absent cytotoxicity. By means of the SEM images produced, it was found that there was adhesion and proliferation of cells on the materials surfaces in the two periods. Therefore, it can be stated that the hydroxyapatite ceramic was presented as a biocompatible material.

A perda do osso alveolar como resultado de doença periodontal ou secundariamente à cirurgia são fontes de inúmeras complicações para os cirurgiões dentistas, e a solução para tal problema se baseiam em procedimentos regenerativos, que é realizada através de enxertos ósseos que estabelecem um arcabouço para a reparação tecidual, aumentando os tecidos ósseos nos defeitos resultantes de trauma ou cirurgias, preenchendo os alvéolos após extração para preservar a altura e espessura do rebordo alveolar e aumentando e/ou reconstruindo o rebordo alveolar. Este projeto tem como objetivo determinar a resistência à tração do osso neoformado com a utilização de diferentes biomateriais. Foram utilizados 60 animais divididos em 3 grupos. O grupo 1A (n=10): GenO; Grupo 1B (n=10): GenMix; Grupo1C (n=10): Bio-Oss; Grupo 2a (n=10): PerioGlas; Grupo 2b (n=10): Polímero de mamona e Grupo 3 (n=10): coágulo. Após 3 meses de nascimento os animais foram submetidos a anestesia intraperitonial e confecção de defeito não critico (3mm de diâmetro) na calvária dos animais, que foram preenchidos com os biomateriais correspondentes a cada grupo. Após 6 meses da cirurgia foi realizada a morte dos animais, os espécimes foram coletados e submetidos ao teste em uma máquina de teste universal. Observa-se que o grupo que obteve maior média de resistência à tração foi o grupo de polímero de mamona (10,932±4,529MPa). Consequentemente, obteve-se uma comparação descrescente na seguinte ordem: Coágulo (9,563±3,74MPa), Bio-Oss (8,706±4,087MPa), GenMix (8,587±3,602MPa), GenOx (7,709±2,416MPa), PerioGlas (7,185±6,837MPa), não notando-se diferença estatisticamente significante entre os grupos. Nas microscopias eletrônicas de varredura na maioria dos grupos observou-se presença de fibras colágenas ao redor do tecido ósseo neoformado e dos remanescentes de enxerto. Conclui-se que o material foi indiferente para o resultado final da resistência à tração, demonstrando uma boa qualidade dos materiais utilizados, tanto quando comparamos os produtos quanto a sua origem (xenogênica, polimérica ou coágulo), ou a sua procedência de fabricação (nacional ou importado). / The loss of alveolar bone as a result of periodontal disease or secondary to surgery are numerous sources of complications for the dentists, and the solution to this problem are based on regenerative procedures, which is conducted through bone grafts that provide a scaffold for tissue repair by increasing the bone tissues in the defects resulting from trauma or surgery, filling the alveoli after extraction to preserve the height and thickness and increasing the alveolar ridge and/or reconstructing the alveolar ridge. This project aims to determine the tensile strength of newly formed bone with the use of different biomaterials. We used 60 animals divided into three groups. Group 1A (n = 10): GenOx; Group 1B (n = 10): GenMix; Group 1C (n = 10): Bio-Oss; Groups 2a (n = 10): PerioGlas; Group 2b (n = 10): castor bean polymer; Group 3 (n = 10): clot. After 3 months of birth the animals were anesthetized intraperitoneally and making non critical defect (3mm diameter) in the calvaria of the animals, which were filled with biomaterials for each group. After 6 months of surgery was performed the animals\' death, the specimens were collected and tested in a universal testing machine. It is observed that the group had a higher average tensile strength was the castor oil polymer group (10.932 ± 4.529 MPa). Consequently, we obtained a comparison descended in the following order: Clot (9.563 ± 3.74 MPa), Bio-Oss (8.706 ± 4.087 MPa), GenMix (8.587 ± 3.602 MPa), GenOx (7.709 ± 2.416 MPa), PerioGlas (7.185 ± 6.837 MPa), not noticing a significant difference between the groups. In scanning electron microscopies in most groups there was presence of collagen fibers around the newly formed bone and remaining graft. It is concluded that the material was irrelevant to the final result of the tensile strength, showing a good quality of materials used, both when comparing products as their source (xenogenic, polymeric or clot), or its origin of manufacture (national or imported).

A perda do osso alveolar como resultado de doença periodontal ou secundariamente à cirurgia são fontes de inúmeras complicações para os cirurgiões dentistas, e a solução para tal problema se baseiam em procedimentos regenerativos. A regeneração é feita através de enxertos ósseos que estabelecem um arcabouço para a reparação tecidual, aumentando os tecidos ósseos nos defeitos resultantes de trauma ou cirurgias, preenchendo os alvéolos após extração para preservar a altura e espessura do rebordo alveolar e aumentando e/ou reconstruindo o rebordo alveolar. Estes procedimentos são realizados através da utilização de enxertos autógenos, alógenos, aloplásticos ou xenogênicos, tendo cada um suas características e indicações. Com o desenvolvimento dos implantes osseointegrados, viu-se a necessidade de manter uma boa quantidade e qualidade óssea. Este projeto tem como objetivo avaliar a resistência a tração e módulo de elasticidade do osso mineralizado e desmineralizado de calvária de ratos, a fim de determinar padrões iniciais (máximos e mínimos) para comparações com possíveis biomateriais de preenchimento de osteoindução e osteocondução. Foram utilizados 24 animais divididos em 2 grupos. O grupo 1 (n=12): módulo de elasticidade sendo que utilizaremos as duas parietais do animal, possibilitando 24 espécimes, subdividido em mineralizado (n=12) e desmineralizado (n=12). O grupo 2 (n=12): microtração, sendo que 12 hemicalvária foram utilizado para o sub-grupo desmineralizado e as outras 12 hemicalvária para o sub-grupo mineralizado. Após a morte dos animais, os espécimes serão coletados e tratados de acordo com o subgrupo (mineralizado e desmineralizado). Os testes foram realizados numa máquina de teste universal (Vitrodyme, V1000). As médias de resistência a tração do mineralizado foi 129,814 ± 34,921 MPa e o desmineralizado foi 18,547 ± 3,682 MPa. Para o módulo de elasticidade obtivemos os valores de 1377,792 ± 208,331 MPa para o grupo mineralizado e 49,669 ± 11,204 MPa para o grupo desmineralizado, ao compararmos o módulo de elasticidade e resistência a tração entre os subgrupo desmineralizado e mineralizado observa-se uma diferença estatisticamente significante para o grupo mineralizado (p<0,001). Conclui-se que o teste de microtração pode ser utilizado para avaliar as propriedades mecânicas nesse modelo experimental biológico. / The loss of the alveolar bone as resulted of periodontal illness or second to the surgery is sources of innumerable complications for the surgeons dentists, and the solution for such problem if it bases on regenerative procedures. Regeneration is made through bone grafts that establish one scaffold for the tecidual repairing, increasing bone fabrics in the resultant defects of trauma or surgeries, filling the alveolous after extraction to preserve the height and thickness of the alveolar rigde and being increased and/or reconstructing the alveolar rigde. These procedures are carried through the use of autogenous, alografts, aloplastic or xenogenics grafts, having each one its characteristics and indications. With the development of the osseointegrated implants, it was seen necessity to keep a good amount and bone quality. This project has as objective to evaluate the resistance the traction and modulus of elasticity of the mineralized bone and desmineralized.of calvarium of rats, similar to determine initial standards (maximum and minimum) for comparisons with possible biomateriais of fulfilling of osteoinduction and osteoconduction. 24 animals divided in 2 groups had been used. Group 1 (n=12): modulus of elasticity being that we will use the two parietals of the animal, having made possible 24 specimens, subdivided in mineralized (n=12) and desmineralized (n=12). Group 2 (n=12): microtraction, being that 12 hemi-calvarium had been used for 12 the desmineralized sub-group and the others hemi-calvarium for the mineralized sub-group. After the death of the animals, the specimens will be collected and treated in accordance with the sub-group (mineralized and desmineralized). The tests had been carried through in a machine of universal test (Vitrodyme, V1000). The resistance averages the traction of the mineralized one was mineralized 129,814 ± 34.921 MPa and the desmineralized one was 18,547 ± 3,682 MPa. For the modulus of elasticity we got values of 1377,792 ± 208.331 MPa for the mineralized group and 49,669 ± 11,204 MPa for the desmineralized group, when comparing the resistance and modulus of elasticity the traction between the desmineralized and mineralized sub-group observes a estatistical significant difference for the mineralized group (p<0,001). Concluded that the microtraction test can be used to evaluate the mechanical properties in this biological experimental model.

Abstract Nickel-titanium shape memory metal Nitinol (NiTi) is a new kind of implant material, which provides a possibility to prepare functional implants activated at body temperature and withstands kinking better than conventional metals. Applications utilizing these unique properties are a target of active research interest. Host reactions to NiTi and to the forces created by functional implants should also be studied. A functional NiTi intramedullary nail, which causes a bending force on the bone, was developed for correcting bone deformities. In the present studies, the action of the device was inverted to induce a bone deformity instead of correcting one, in order to test the hypothesis that bone modelling can be controlled using such functional nail. Implanting the nail into the medullary cavity of rat femur for twelve weeks caused bowing of the bone, retardation of its longitudinal growth, and thickening of the bone and the cortex. In another study the effects of functional and straight nails were compared. Bowing of the bone and significant overall thickening of the bone and the cortex were associated only with the functional nail, while the straight nail induced only minor thickening of the bone. Retardation of longitudinal growth was seen in both groups, and this may have been caused by perforating the distal epiphyseal plate during the nailing. Finite element model of the bone-nail combination was also created. Porous NiTi was studied as a bone graft substitute by filling a bone defect in the distal femoral metaphysis of a rat bone with porous NiTi implants of different porosities. After 30 weeks, porosity of 66.1% (mean pore size (MPS) 259μm) showed the best bone-implant contact (51%). However, porosity of 46.6% (MPS 505μm) with 39% bone-implant contact was not significantly inferior in this respect and showed a significantly lower incidence of fibrosis within the implant and thus seemed to be the best choice for a bone graft substitute, out of the porosities tested here. The porosity of 59.2% (MPS 272μm) showed lower contact values. NiTi tendon suture material was studied by implanting NiTi sutures into rabbit tendon and subcutaneous tissues for two, six, and twelve weeks. NiTi proved to be stronger than polyester, which served as control material. The encapsulating membrane was minimal with both materials, suggesting good biocompatibility in tendon tissue. The implantation did not affect the strength properties of either material. On the basis of the present studies, NiTi provides a possibility to develop new kinds of implants for correcting bone deformities, for filling bone defects in weight-bearing locations and a good candidate for a tendon suture material.

Abstract Reindeer antler was studied for its possible use as a bone implant material. A molecular biological study showed that antler contains a growth factor promoting bone formation. Ectopic bone formation assay showed that antler is not an equally effective inducer as allogenic material. Ectopic bone formation assay was optimised for biocompatibility studies of orthopaedic NiTi implants. Ti-6Al-4V and stainless steel were used as reference materials. The assay showed differences in bone mineral densities, with superior qualities in NiTi. The rate of endochondral ossification varied between the implants, NiTi ossicles had larger cartilage and bone areas than ossicles of the two other materials. The cytocompatibility of NiTi was studied with three different methods. Cell viability, cell adhesion and TGF-β1 concentration were assessed in ROS-17/2.8 cell cultures. Cells grown on NiTi had better viability than cells grown on pure nickel or stainless steel. Cell attachment on the materials was studied with paxillin staining of focal contacts. The number of focal contacts was clearly higher in cells grown on NiTi than in cells grown on pure titanium, pure nickel or stainless steel. TGF-β1 concentration was measured with ELISA. The results showed that there was only some minor variation between NiTi, pure titanium and stainless steel. Nickel showed a lower TGF-β1 concentration. Taken together, these results suggest that NiTi is well tolerated by ROS-17/2.8 cells. The cytocompatibility of stainless steel is not so good as that of NiTi. The same tests were used to study the effects of the surface roughness of the implant on cytocompatibility. Three different surface roughness grades were compared in cell cultures on NiTi and titanium alloy discs. Titanium alloy was subjected to two different heat treatments, to compare the effects of the treatments on cytocompatibility. The studies showed that NiTi had a lesser impact on cell viability and attachment than titanium alloy. Further, rough NiTi was found to be a better tolerated surface than the others. In this study, heat treatment of titanium alloy at +850° C did not interfere with cell viability or attachment, as did the +1050° C treatment of the alloy. On the contrary, TGF-β1 concentrations decreased on the +850° C treated alloy and were approximately same on the +1050° C treated alloy and on NiTi.

Abstract The improvement of implant biocompatibility is constantly under investigation. Titanium is a standard biomaterial that performs well in dental and orthopedic implantations. However, detrimental adverse effects resulting from e.g. biomaterial properties, inflammatory responses and surgical procedures occasionally occur. Coating the biomaterials aims at increasing the proportion of successful operations. Pectins, large plant cell wall polysaccharides, are innovative, modifiable, and potentially anti-inflammatory candidates for biomaterial nanocoatings. In this thesis, covalently-grafted pectin fragments (modified hairy regions, MHRs) modified either in vitro (from apple) or in vivo (from potato) were tested. Cell culture vessels and titanium substratum coated with the apple-MHRs, MHR-A and a further-tailored fragment type, MHR-B, were compared with controls for their ability to support proliferation and differentiation of osteoclasts and osteoblasts. Cells grew and differentiated on MHR-B and on the control surfaces; MHR-A did not perform well in these assays. Genetically-engineered potato MHRs did not support bone cell growth to the same extent as apple MHR-B, but nonetheless the possibility to manipulate cellular proliferation with specific in vivo – modifications of pectins was introduced. When implanted into rat soft tissues, neither of the apple MHRs provoked severe acute inflammatory reactions, which indicates good in vivo - tolerance of these botanical macromolecules. These studies illustrate the biocompatibility of MHRs, and the directions towards which they could be further tailored. In terms of clinical use, their tolerability in vivo is especially significant.