L’hydroxyapatite phosphocalcique (HA) est largement utilisée en tant que substitut osseux. Sa composition chimique est proche de celle du minéral osseux ce qui en fait une céramique ostéoconductrice. Les hydroxyapatites silicatées (SiHA) sont actuellement étudiées. La présence de silicium au sein de la structure apatite pourrait accroître la bioactivité des implants. Cependant, ces substituts ne sont pas ostéoinducteurs. Ils n'ont pas la capacité d'induire la formation de tissus osseux, ce qui limite leur utilisation en chirurgie réparatrice. L'une des stratégies envisagées afin de les rendre ostéoinducteurs est de fonctionnaliser leur surface par des molécules biologiquement actives via des organosilanes. L’objectif de ce travail est d'étudier l’influence de l’incorporation du silicium dans l’hydroxyapatite et de la fonctionnalité (i.e. nombre de groupements hydrolysables) des amino-éthoxy-silanes sur le mode de greffage et la quantité de molécules à la surface des substrats. Les céramiques HA et SiHA silanisées ont été élaborés et caractérisées par spectroscopie de photoélectrons X et par analyse thermogravimétrique couplée à la spectrométrie de la masse. Les résultats ont confirmé un greffage covalent quelles que soient la nature du substrat et la fonctionnalité de l'organosilane. La quantité greffée augmente avec la fonctionnalité de l’organosilane. La présence de silicium au sein du substrat favorise le nombre de chaines éthoxyles impliquées dans la réaction d’hétérocondensation et diminue la quantité d’organosilanes greffée. Les organosilanes sont répartis d’une façon non homogène sur la surface mais sans formation d’une couche recouvrant celle-ci en totalité. / Calcium phosphate hydroxyapatite (HA) is widely used as bone substitute. Its chemical composition is close to the mineral part of bone which induces osteoconductive ceramics. Silicated hydroxyapatites (SiHA) are being studied. The incorporation of silicon in the hydroxyapatite structure would increase the bioactivity of the implant. However, these substitutes are not osteoinductive. They do not have the ability to induce bone formation which limits their use in reparative surgery. In order to obtain osteoinductive ceramics, it is possible to functionalize their surface by biologically active molecules via organosilanes. The present work studies the influence of silicon incorporation in the hydroxyapatite structure and the functionality (i.e. number of hydrolysable groups) of amino-ethoxy-silanes on the grafting mode and the quantity of molecules at the substrates surface. HA and SiHA silanised ceramics have been produced and characterized by means of X-ray photoelectron spectroscopy and thermogravimetric analysis coupled with mass spectrometry. The results confirmed the covalent grafting whatever the nature of the substrate and the functionality of the organosilane might be. The grafted amount increases with the functionality of organosilane. Moreover, the presence of silicon within the substrate promotes the number of ethoxyl chains involved in the heterocondensation reaction and decreases the grafted amount of organosilanes. The organosilanes are not homogeneously distributed on the surface but without formation of a layer covering the entire surface.

En France, environ 9,8% de la population souffre de lithiase urinaire. Nous proposons, ici, une nouvelle approche afin de caractériser ces matériaux et d'obtenir une meilleure compréhension de leur formation. Les médecins utilisent principalement des techniques FTIR pour déterminer les principales phases présentes dans une calcification pathologique donnée. Les méthodes de RMN semblent appropriées pour déterminer plus précisément la composition de ces calcifications pathologiques. Très peu d'analyses RMN de calculs rénaux et d'analogues synthétiques ont été réalisées à ce jour.Premièrement, les trois phases d'oxalate de calcium ont été synthétisées (CaC2O4?nH2O avec n = 1, 2, 3) en mettant en ¿uvre des protocoles originaux. L'analyse des spectres 1H, 13C CPMAS et 43Ca MAS à ultra-haut champ magnétique en abondance naturelle permet une identification précise des différentes phases et peut être interprétée en tenant compte du nombre d'inéquivalents des sites cristallographiques. L'hydroxyapatite a été étudiée en parallèle car pouvant également apparaître lors de lithiase urinaire.Enfin, nous montrons les résultats obtenus à partir de DNP MAS à 100 K sur les échantillons synthétiques. L'affinité de la combinaison biradical/solvant reste un facteur limitant pour les matériaux hétérogènes tels que ceux qui sont analysés dans cette étude. Nous devons être en mesure de trouver une combinaison appropriée pour les matériaux multi-composants complexes afin d'obtenir un gain équivalent sur tous les signaux. Finalement, il est démontré qu'il est possible de transposer l'approche méthodologique présentée ci-dessus à l'étude des calculs rénaux. / In France, about 9.8% of the population suffer from urolithiasis. The treatment of kidney stones, composed of 72% of hydrated calcium oxalates (CaC2O4∙nH2O) with n = 1, 2, 3, represents almost 900 million euros in annual spending and it is therefore necessary to understand the in vivo formation of these stones. Here we propose a new approach in order to characterize kidney stones and have a greater understanding of their formation. Physicians primarily use FTIR techniques to determine the major phases present in a given pathological calcification. NMR methods appears suitable to determine more accurately the composition of these pathological calcifications. Very few NMR analyzes of kidney stones and synthetic analogues were conducted to date. In a first step, the three phases of calcium oxalate were synthesized by implementing original protocols. Analysis of the 1H, 13C CP MAS and MAS 43Ca ultra-high magnetic field spectra at natural abundance allow precise identification of the different phases and can be interpreted by taking into account the number of inequivalent crystallographic sites. The hydroxyapatite was studied in parallel as it may also appear in urolithiase. Finally, we show results obtained from DNP MAS at 100 K on the synthetic samples. The affinity of the biradical/solvent combination remains a limiting factor for heterogeneous materials such as those analyzed in this study. We have to be able to find a suitable combination for complex multi-component materials and to obtain an equivalent gain on all signals. In a last step, it is demonstrated that it is possible to transpose the methodological approach presented above to the study of kidney stones.

Dans ce travail, nous avons élaboré, par voie sol-gel, un nouveau verre bioactif mésoporeux : 92S6 (92% SiO2, 6% CaO, et 2% P2O5). Cette synthèse a été développée en variant les paramètres telles que la température, la nature du tensioactif (ionique (le CTAB) ou non ionique (le P123)) et la longueur de la chaîne carbonée pour chaque type de tensioactif et en étudiant leurs effets sur les propriétés texturales du verre. Des études physico-chimiques et biologiques in vitro ont été menées sur ces biomatériaux. Par la suite, nous avons réalisé une étude structurale de l'eau confinée dans les pores du verre synthétisé qui a révélé une modification de l'ordre locale en comparaison avec l'état massique (bulk). Ceci permet un contrôle de la quantité de principe actif à incorporer dans les pores, du taux d'adsorption et de la cinétique de libération. La diffraction des Rayons X aux petits angles, l'analyse texturale par BET et les images de la microscopie électronique en transmission ont mis en évidence l'influence des paramètres de synthèse sur les propriétés texturales des verres synthétisés. En effet, l'échantillon synthétisé avec le tensioactif ionique (CTAB) présente une structure mésoporeuse non ordonné, alors que l'utilisation du tensioactif non-ionique (P123) favorise l'obtention d'un verre mésoporeux ayant une structure poreuse ordonnée. Dans le cas des verres synthétisés avec les différentes longueurs de la chaîne carbonée des tensioactifs, nous avons démontré que la longueur de la chaîne alkyle du tensioactif ionique détermine la morphologie des particules et les caractéristiques texturales des 3 verres, alors que nous avons noté une absence de différences significatives dans la texture des matériaux dans le cas des 3 verres synthétisés avec les tensioactifs non ioniques. Nous avons aussi mis en évidence l'influence de la température de synthèse sur les propriétés texturales des verres synthétisés. Nous avons démontré que la température de synthèse agit sur le diamètre des pores de manière différente selon la nature du tensioactif. L'augmentation de la température de synthèse conduit à une augmentation de la dimension des cavités dans le cas des verres synthétisés avec le P123, alors que dans le cas des verres synthétisés avec le tensioactif ionique CTAB, l'augmentation de la température de synthèse conduit à une diminution des diamètres des pores. Nous avons démontré que la morphologie des particules et les caractéristiques texturales des verres influent sur la bioactivité de ces derniers. Nous avons obtenu des verres bioactifs comme en témoigne la formation de la couche d'hydroxyapatite à leurs surfaces une fois immergés dans le SBF, ce qui explique l'intérêt des ces verres dans le domaine de la chirurgie orthopédique. / In this study, a new mesoporous bioactive glass 92S6 (92% SiO2, 6% CaO, and 2% P2O5), was prepared by sol-gel method. The purpose of this study was first to investigate the effect of synthesis parameters such as aging temperature, surfactant type and surfactant carbon chain length on the glass textural properties and then to investigate these effect on the glass bioactivity. Then, we conducted a structural study of water confined in the pores of the synthesized glass. This study revealed a change of the local order of confined water in comparison with the bulk. This allows control of drugs to be incorporated into the pores, the adsorption rate and the release kinetics. The small-angle XRD patterns (SAXRD), the Brunauer–Emmett–Teller (BET) method and the Transmission Electron Microscopy (TEM) studies evidence the influence of the structure-directing agent (ionic surfactant CTAB or non-ionic P123) in the generation of unorganized or well ordered pores in the 92S6 glass. We also demonstrated that when we synthesized glasses with a same composition using the same technique (sol-gel), in presence of surfactants with different carbon chain length, there is a direct correlation between the surfactant and textural properties and structural characteristics. As observed from small-angle XRD patterns and TEM images, synthesis at different temperatures lead to the formation of mesoporous glasses. The properties of these glasses are extremely dependent on the synthesis temperature. We demonstrated also that the bioactivity may be significantly modified by a change on glass textural properties. Finally, the samples were analyzed to quantify the apatite formation ability when soaked in SBF solution. The physico-chemical studies showed apatite-like phases formed at the glasses surface when soaked in SBF. So, our mesoporous glasses are bioactive. These characteristic mesoporous glasses will display good future applications in bone tissue repairing and engineering.

published_or_final_version / abstract / Mechanical Engineering / Doctoral / Doctor of Philosophy

Dental restorative cements are placed in a harsh oral environment where they are subjected to thermal shock, chemical degradation, and repeating masticatory force. The ideal restorative dental cements should have superior mechanical properties, chemical stability, aesthetic, good handling properties, biocompatibility, antibacterial properties, and preferably bioactivity. This thesis presents research on dental restorative cements with enhanced properties. The overall aim was to increase the bioactivity and antibacterial properties of dental restorative cements without affecting their other properties. The effect from adding calcium silicate to glass ionomer cement (GIC) was investigated. The results showed that calcium silicate could increase the bioactivity and reduce the cytotoxicity of conventional glass ionomer cement without compromising its setting and mechanical properties. Hydroxyapatite (HA) with a high aspect ratio and thin nacreous-layered monetite sheets were also synthesized. Nano HA particles with an aspect ratio of 50 can be synthesized by both precipitation and hydrothermal methods. The aspect ratio was controlled via the pH of reaction medium. Thin nacreous-layered monetite sheets were synthesized through a self-assembly process in the presence of an amine based cationic quaternary surfactant. Temperature, pH, and presence of surfactant played essential roles in forming the nacreous-layered monetite sheets. Then the effect from adding silver doped HA and monetite particles was investigated. The results showed that the antibacterial properties of GIC could be increased by incorporating silver doped HA and monetite particles. Further examination showed that the pH change, F- ion release, and concentration of released Ag+ ions were not responsible for the improved antibacterial properties. The quasi-static strengths and compressive fatigue limits of four types of the most commonly used dental restorations were evaluated. In our study, resin modified GIC and resin-based composite showed superior static compressive strength and fatigue limits compared to conventional GIC. The static compressive strength of dental cements increased with the aging time. However, aging had no effect on the compressive fatigue limit of resin modified GIC and resin-based composite. The compressive fatigue limit of conventional GIC even showed a drastic decrease after aging.

Abstract Objective Cranioplasty is routinely performed in neurosurgery. One of its underestimated problems is the high postoperative complication rate of up to 40%. Due to the lack of good prospective studies and the low number of patients (5–20 each year) receiving alloplastic materials, decisions in favor or against a certain material are based on subjective empirical or eco-nomic reasons. The main goal of this first prospective, randomized multicenter study in Germany of custom-made titanium and hydroxyapatite (HA) implants was to compare lo-cal and systemic infections related to the implant within the first six months after implanta-tion. Secondary objectives included comparing reoperation rates, the complication rate, clinical and neurological outcome, and health-related quality of life. Methods The study included patient screening and randomization 6–8 weeks before operation, pre-, intra and postoperative documentation until discharge, and postoperative follow-ups after one and six months. Approval for the study was obtained from the local ethics committee. The study design was published on www.clinicaltrials.gov. Results A total of 52 patients were included in the study. The rate of local, implant associated wound infection in the HA group was 2 out of 26 (7.7%) patients and 5 out of 24 (20.8%) patients in the titanium group (p=0.407 n.s.). Systemic inflammation within six months after operation affected none of the patients in the HA group and 4 out of 24 (37.5%) pa-tients in the titanium group (p=0.107 n.s.). In both groups, 7 patients had to be reoperated after the six-month follow-up (26.9% in the HA group and 29.2% in the titanium group; n.s.). Re-surgery with explantation was necessary in 3 patients in each group (11.5% in the HA group and 12.5% in the titanium group; n.s.). The results demonstrated a significantly higher number of epidural hematomas in the HA group compared to none in the titanium group. Altogether, 46 AE were measured in 27 patients (54%). An improvement of the neurologi-cal outcome after six months was experienced by 43% of the patients in the HA group and 26.3% in the titanium group (p=0.709 n.s.). Conclusion The study emphasizes that cranioplasty is a high-risk intervention. Compared to tita-nium, HA shows a benefit in the infection rate and the neurological outcome, but has at the same time a higher postoperative risk for epidural hematoma. Depending on the individual conditions both materials have their place in future therapy of cranioplasty.

O objetivo deste trabalho foi produzir hidroxiapatita (HA) contendo diferentes tipos de própolis de origem brasileira e avaliar a possível atividade antimicrobiana dos materiais. Os extratos etanólicos de própolis (EEP) vermelha, verde e marrom foram obtidos em solução alcoólica 80%. EEP verde e vermelha (8 mg/mL e 20 mg/mL) foram incorporados ao material a 10% (m/v) via atomização (spray drying), obtendo-se HA-GP8, HA-GP20, HA-RP8 e HA-RP20. Os EEP e os materiais foram caracterizados quanto ao conteúdo de compostos fenólicos e flavonoides totais. A atividade antimicrobiana dos EEP foi avaliada por difusão em ágar, concentração inibitória mínima (CIM) e bactericida mínima (CBM) frente à Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis) e Escherichia coli (E. coli). Os pós de HA incorporados com própolis foram avaliados frente à S. aureus por contagem de colônias bacterianas em placa, CIM e CBM. A caracterização dos materiais foi realizada por difração de raios X (DRX), espectroscopia na região do infravermelho com transformada de Fourier (FTIR) e microscopia eletrônica de varredura (MEV). A citotoxicidade foi determinada pelo cálculo da viabilidade celular, realizada pelo método de incorporação do vermelho neutro. Para os extratos, o conteúdo de fenólicos variou entre 278,3 e 325,6 mg EAG/g ES. EEP vermelha mostrou maior conteúdo de flavonoides (112,4 mg EQ/g ES) em relação aos outros EEP (48,4 e 52,3 mg EQ/g ES), apresentando maior atividade inibitória (CIM) frente à S. aureus e S. Epidermidis (12,5 µg/mL), comparado ao EEP verde (100 e 200 µg/mL) e marrom (200 µg/mL). Para CBM, os EEP vermelha e verde foram mais efetivos (800 µg/mL) comparado ao EEP marrom (1600 µg/mL), frente às mesmas bactérias. Entretanto, não foi observada atividade frente à E coli. A caracterização dos pós incorporados ou não com própolis apresentou estrutura cristalina e morfologia aparentemente esférica, indicando diminuição no grau de aglomeração com a adição de própolis. FTIR indicou a presença de grupos funcionais característicos para HA e própolis. Os materiais apresentaram alta liberação de fenólicos (228,3 a 327,6 mg EAG/g ES) e menores quantidades de flavonoides (14,0 a 35,8 mg EQ/g ES), sendo as maiores quantidades de flavonoides atribuída à HA contendo própolis vermelha. Foi verificado efeito bactericida a partir de 0,5 h (HA-RP20 e HA-GP20) e 1 h (HA-RP8 e HA-GP8) e menor atividade inibitória (CIM) para HA-GP20 e HA-GP8 (175,4 e 182,0 µg/mL) e para HA-RP20 e HA-RP8 (51,7 e 66,8 µg/mL), comparado aos EEP. Entretanto, observou-se maior atividade bactericida (CBM) para HA-GP20 e HA-GP8 (701,5 e 728,0 µg/mL) e para HA-RP20 e HA-RP8 (206,5 e 267,0 µg/mL), em relação aos EEP. O ensaio de citotoxicidade mostrou valores para IC50 (concentração que reflete 50% da viabilidade celular) de 387,1 e 84,8 µg/mL para as amostras HA-GP8 e HA-RP8, respectivamente. Considerando os resultados obtidos neste trabalho, sugere-se que a HA incorporada com própolis (HA-GP8 e HA-RP8) pode ser utilizada como possível agente antimicrobiano, inibindo o crescimento de S. aureus, respeitando-se os valores máximos para IC50. Entretanto, não poderia ser utilizada como agente bactericida, uma vez que nestas condições os materiais apresentaram efeito citotóxico. / The aim of this study was to produce hydroxyapatite (HA) containing different types of propolis of Brazilian origin and to evaluate the possible antimicrobial activity materials. The ethanolic extracts of red, green and brown propolis (EEP) were obtained in alcoholic solution 80%. Green and red EEP (8 mg / mL and 20 mg / mL) were incorporated into the 10% (m/v) via atomization (spray drying), obtaining HA-GP8, HA-GP20, HA-RP8 and HA -RP20. EEP and materials were characterized regarding the content of phenolic compounds and total flavonoids. The antimicrobial activity of the EPS was evaluated by diffusion in agar, minimum inhibitory concentration (MIC) and minimum bactericidal (MBC) against Staphylococcus aureus (S. aureus), Staphylococcus epidermidis (S. epidermidis) and Escherichia coli (E. coli). The HA powders incorporated with propolis were evaluated against S. aureus by plate colony counting , CIM and CBM. Materials characterization was made by X-ray diffraction (XRD), Fourier Transform Infrared Spectroscopy (FTIR) and Scanning Electron Microscopy (SEM). The cytotoxicity of the materials was determined by cell viability calculation, which was made by the neutral red incorporation method. For extracts, phenolic content ranged from 278.3 to 325.6 mg EAG/g ES. The red EEP showed a higher flavonoid content (112.4 mg EQ/g ES) than the other EEP (48.4 and 52.3 mg EQ/g ES), showing a higher inhibitory activity (MIC) against S. aureus and S. epidermidis (12.5 µg/mL) compared to green EEP (100 and 200 µg/mL) and brown (200 µg/mL). For CBM, red and green EEP were more effective (800 µg/mL) compared to brown EEP (1600 µg/mL) against the same bacteria. However, no activity was observed against E coli. The characterization of the powders incorporated or not with propolis presented crystalline structure and apparently spherical morphology, indicating a decrease in the degree of agglomeration with the addition of propolis. FTIR indicated the presence of functional groups characteristic for HA and propolis. The materials presented high phenolic release (228.3 to 327.6 mg EAG/g ES) and lower amounts of flavonoids (14.0 to 35.8 mg EQ/g ES), with the highest amounts of flavonoids attributed to HA containing red propolis. The bactericidal effect for all materials was observed within the interval of 0.5 (HA-RP20 and HA-GP20) to 1 hour (HA-RP8 and HA-GP8). The materials showed lower inhibitory activity (MIC) for HA-GP20 and HA-GP8 (175, 4 and 182.0 µg/mL) and for HA-RP20 and HA-RP8 (51.7 and 66.8 µg/mL) compared to EEP. However, higher bactericidal activity (MBC) was observed for HA-GP20 and HA-GP8 (701.5 and 728.0 µg/mL) and for HA-RP20 and HA-RP8 (206.5 and 267.0 µg/mL) when compared to the EEP. The cytotoxicity assay showed values for IC50 (concentration that reflects 50% of cellular viability) of 387.1 and 84.8 µg/mL for HA-GP8 and HA-RP8 samples, respectively. Considering the results obtained in this work, it is suggested that HA incorporated with propolis (HA-GP8 and HA-RP8) can be used as a possible antimicrobial agent, inhibiting the growth of S. aureus, respecting the maximum values for IC50. However, it could not be used as a bactericidal agent, since under these conditions the materials had a cytotoxic effect.

Um dos grandes desafios para o tratamento de defeitos ósseos extensos na região bucomaxilofacial têm sido o desenvolvimento de um biomaterial substituto ósseo ao enxerto autógeno. No presente trabalho avaliou-se a formação óssea e a biodegrabilidade do osso desproteinizado bovino Bio-Oss® e do seu similar GenOx Inorg® e da cerâmica bifásica GenPhos® XP no processo de reparo de defeitos ósseos cranianos em coelhos, comparativamente ao osso autógeno (controle positivo) e coágulo sanguíneo (controle negativo). Foram realizados cirurgicamente defeitos bilaterais de 8-mm de diâmetro nos ossos parietais de 39 coelhos. A seguir os defeitos foram preenchidos aleatoriamente com 0,1cm3 de material ou coágulo conforme cada grupo de tratamento. Após os períodos de 4, 8 e 24 semanas os crânios foram coletados, analisados no microtomógrafo e processados histologicamente. O percentual de volume do defeito ocupado pelo material e osso neoformado foi avaliado pela microtomografia e histomorfometria, enquanto que, para a medula óssea, tegumento e tecido conjuntivo, apenas pela análise histomorfométrica. Os resultados quantitativos obtidos foram comparados estatisticamente pela ANOVA a dois critérios (período e tratamento) e teste de Tukey com p<0,05. A intensidade da associação linear dos dados microtomográficos e histomorfométricos avaliada pelo coeficiente de correlação de Pearson, mostraram correlação moderada a forte. Nos períodos iniciais de reparo (30 e 60 dias), os defeitos tratados com Bio-Oss®, GenOx® Inorg e GenPhos® XP apresentaram manutenção do volume do material enxertado (Vvi médio de 34% ) e formação óssea menor e mais imatura em relação grupo autógeno (Vvi = 22% vs. 32% no grupo autógeno). No período mais tardio (180 dias) a quantidade de formação óssea foi estatisticamente similar nos grupos Bio-Oss® (Vvi = 27%), GenOx® Inorg (Vvi = 26%) e GenPhos® XP (Vvi = 20%) porém, o GenOx® Inorg promoveu a formação de um tecido ósseo mais organizado e com maior acúmulo de biomaterial+osso+medula óssea (Vvi = 67,9%) comparado ao GenPhos® XP (Vvi =58,9%) e Bio Oss (Vvi = 55,6%) mas, inferior ao do enxerto autógeno (Vvi = 78%). Os resultados aqui obtidos permitem concluir que o osso autógeno promove rápida formação e maturação óssea, porém não consegue promover o reestabelecimento completo da díploe removida cirurgicamente. Os materiais BioOss, GenOx® Inorg e GenPhos® XP são excelentes materiais osteocondutores levando a formação óssea em toda extensão do defeito, sendo o GenOx® Inorg o que apresenta menor grau de reabsorção e maior e melhor preenchimento do defeito. / One major challenge for treatment of critical size defects in maxillofacial region has been the development of a substitute biomaterial to the autogenous bone grafts. In present study we evaluated the bone formation and biodegradability of deproteinized bovine bone Bio-Oss® and GenOx® Inorg, and biphasic calcium phosphate GenPhos XP® during bone repair process in rabbits cranial defects compared to autogenous bone (positive control) and blood clot (negative control). In parietal bone of 39 rabbits were made bilateral 8-mm diameter defects, which were filled randomly with 0,1cm3 material or clot as each treatment group. After periods of 4, 8 and 24 weeks skulls of animals were collected, analyzed the MicroCT scanner and histologically processed. The percentage of defect volume occupied by biomaterial and new-formed bone were assessed by histomorphometry and microtomography, while the bone marrow, connective tissue and tegument only by first analysis. The quantitative data were compared by two-way ANOVA analysis (time and treatment) and Tukey\'s test at p <0.05. The intensity of the linear association of MicroCT and morphometric data evaluated by the Pearson correlation coefficient, showed moderate to strong correlation. In the early repair periods (30 and 60 days), the defects treated with Bio- Oss, GenOx® Inorg and GenPhos® XP showed maintenance of the graft material volume (average Vvi of 34%) and lower and more immature bone compared autograft group (Vvi = 22% vs. 32% in the autograft group). In the later period (180 days) the amount of bone formation was statistically similar to the groups Bio-Oss® (Vvi = 27 %), GenOx® Inorg (Vvi = 26%) and GenPhos® XP (Vvi = 20%) however, the bone formation in GenOx® Inorg was more organized and with greater accumulation of particles + bone tissue + bone marrow (Vvi = 67.9%), when compared to GenPhos® XP (Vvi = 58.9%) and Bio-Oss® (Vvi = 55.6%) but lower than the autograft (Vvi = 78%). It was concluded that the autogenous bone promotes rapid bone formation and maturation, but cannot promote the complete reestablishment of diploe surgically removed. The Bio-Oss®, GenOx® Inorg and GenPhos® XP are excellent osteoconductive materials leading to bone formation in the full extent of the defects, and the GenOx® Inorg showing less absorption promotes more and better defect filling.

O intuito do desenvolvimento de biomateriais potencialmente aplicáveis como implantes de substituição óssea é sintetizar superfícies que induzam respostas biológicas positivas quando em contato com o tecido hospedeiro. Para isso, faz-se necessário não somente estudos de composição química, mas também é importante buscar compreender propriedades físicas e estruturais dos sistemas que estes materiais irão substituir. O osso natural é constituído por compostos orgânicos, especialmente proteínas como o colágeno, reforçado com minerais, tais como hidroxiapatita. O processo pelo qual ocorre a formação do componente inorgânico é chamado biomineralização. Uma maneira de introduzir macromoléculas em sistemas modelo para estudos de biomineralização é a formação de monocamadas Langmuir, que são camadas monomoleculares altamente organizadas de moléculas anfifílicas na interface líquido-ar. Essas monocamadas podem ser transferidas para suportes sólidos formando filmes de Langmuir-Blodgett. Neste estudo, foi investigado a incorporação de colágeno tipo-I em monocamadas insolúveis contendo os fosfolipídeos 1,2-dipalmitoil-sn-glicero-3-fosfatidilcolina (DPPC) ou ácido octadecilfosfônico (OPA), bem como a transferência dessas monocamadas para suportes de titânio por meio da técnica de Langmuir-Blodgett. Estes filmes foram utilizados como matrizes para o crescimento de hidroxiapatita biomimética após a exposição dos discos de titânio modificados a uma solução que simula a concentração de íons e pH do plasma sanguíneo humano. As amostras foram caracterizadas por microscopia eletrônica de varredura (MEV), microscopia de força atômica (AFM), espectroscopia vibracional na região do infravermelho (FTIR) e difração de raios-X (DRX). A razão molar Ca/P foi determinada por espectroscopia de dispersão de energia de raios-X (EDX). Propriedades dos biomateriais como molhabilidade, rugosidade e energia livre de superfície também foram estudadas. A biocompatibilidade das amostras foi investigada in vitro utilizando-se ensaios de viabilidade de osteoblastos. O colágeno interage com os fosfolipídios na interface água/ar, como evidenciado pelas isotermas de pressão de superfície, obtidas na presença da proteína. Além disso, a estabilidade das monocamadas de Langmuir foi aumentada devido à presença de colágeno na subfase. A incorporação de colágeno e fosfolipídios em filmes Langmuir-Blodgett sobre as superfícies de titânio são fatores determinantes para a formação de filmes inorgânicos altamente organizado, contínuos e uniformes, compostos por nanopartículas de hidroxiapatita biomimética, semelhante à encontrada no osso natural. As modificações nas superfícies aumentaram a proliferação de osteoblasto no titânio indicando que este material não é tóxico a este tipo de célula. / The development potential of biomaterials for bone replacement is guided by the necessity to provide a surface that induces positive biological responses when the material is in contact with the host tissue. For this, it is necessary the study of the chemistry and surface composition and also to understand the physical properties and the structural organization of the natural system which is in contact with the biomaterial. Natural bone is composed by organic compounds, especially protein like collagen, reinforced with minerals, such as hydroxyapatite. The process by which the formation of the inorganic component takes place is named biomineralization. One way to introduce macromolecules in model systems for biomineralization studies is the formation of Langmuir monolayers, which are highly organized monomolecular layers of amphiphilic molecules adsorved at the liquid-air interface. These monolayers can be transferred to solid substrate to form the Langmuir-Blodgett films. In this study, it was investigated the incorporation of type-I collagen in insoluble Langmuir monolayers containing either 1,2-dipalmitoyl-sn-glycero-3-phosphatidylcholine (DPPC) or octadecylphosphonic acid (OPA) and their transference to titanium supports using the Langmuir-Blodgett technique. These films were used as matrices to the growth of biomimetic hydroxyapatite by the exposure of the modified titanium discs to a solution that mimics the ions concentration and pH of the human blood plasma. The samples were characterized by scanning electron microscopy (SEM), atomic force microscopy (AFM), vibrational spectroscopy in the infrared region (FTIR), and X-ray diffraction (XRD). The Ca/P molar ratio was determined by energy dispersive X-ray spectroscopy (EDS). The properties of the biomaterials as wettability, roughness, and surface free energy were also studied. The biocompatibility of the samples was studied in vitro using osteoblasts viability assays. The collagen interacts with the phospholipids at air/water interface as evidenced by the pressure surface-surface area isotherms obtained in the presence of the protein. Moreover, the stability of the Langmuir monolayers was increased when collagen is present at the subphase. The incorporation of collagen and phospholipids on the titanium surfaces is important to the formation of organized, continuous and uniform inorganic films composed by biomimetic hydroxyapatite nanoparticles, similar to that found in the natural bone. These films are not toxic to osteoblasts as evidenced by the stimulation of their proliferation on the titanium-modified samples.

A hidroxiapatita (HA) é o principal constituinte mineral de ossos e dentes, e têm sido foco de grande interesse na biomedicina devido sua excelente biocompatibilidade e bioatividade, em especial para aplicação em implantes dentários e ortopedia. Na odontologia estudos mostram o sucesso do uso de nanopartículas (NP) de HA para tratamento de hipersensibilidade dentinária, remineralização de lesões de cárie em esmalte, reparação e prevenção de lesões de erosão iniciais. No entanto, existem evidências que o tamanho do cristal desempenha um papel essencial na formação dos tecidos duros e possui significante influência nas propriedades intrínsecas destes cristais, incluindo solubilidade e, consequentemente, biocompatibilidade. Este trabalho teve como objetivo verificar a síntese e caracterização de nanopartículas de hidroxiapatita funcionalizadas e avaliar a influência de sua aplicação no processo adesivo em dentina. Nanopartículas de HA com diferentes tamanhos e morfologia foram sintetizadas pelo método de precipitação em solução aquosa. Foi investigada a influência de dois novos ligantes, ácido deoxicólico (AD) e ácido abiético (AB), no preparo da NP de HA. As NP foram caracterizadas através das análises de espectroscopia por dispersão de raios X (EDX), Difratometria de Raios X (DRX), Espalhamento de Luz Dinâmica (DLS), Microscopia Eletrônica de Transmissão (MET), Microscopia Eletrônica de Varredura (MEV), Microscopia de Força Atômica (AFM). Os resultados mostraram que a adição destes ligantes alteram a morfologia mas não o arranjo cristalino da HA. Para o estudo da interação das NP de HA com o colágeno foi utilizado a NP modificada por AD. Para esta análise da influência da aplicação destas NP durante o procedimento adesivo, foram utilizados dentes bovinos. Os dentes bovinos foram separados em quatro grupos Controle (C), Controle termociclado (CTC), nanopartícula funcionalizada com ácido deoxicólico (AD) e Deoxicólico termociclado (ADTC). Foi realizado o teste de cisalhamento 24 horas após a realização do procedimento adesivo e após a termociclagem (10 000 ciclos). Foram realizadas imagens da interface adesiva antes e depois da termociclagem, através da Microscopia Confocal Raman. Os resultados mostraram um aumento nos valores da resistência de união inicial do grupo AD quando comparados ao grupo C, e a manutenção do valor após a ciclagem térmica no grupo ADTC que foi estatisticamente superior ao grupo CTC (p<0,05). Não foi observada alteração no padrão da camada híbrida formada, mostrando um possível benefício da utilização deste nanomaterial na odontologia. / Hydroxyapatite (HA) is the major mineral component of bones and teeth, and has been the focus of great interest in biomedicine because of its excellent biocompatibility and bioactivity, in particular for application in dental and orthopedic implants. In dentistry studies have shown the successful use of nanoparticles (NP) of HA for the treatment of dentin hypersensitivity, remineralization of caries lesions in enamel, repair and injury prevention of initial erosion. However, there is evidence that the size of the crystal plays an essential role in the formation of hard tissues and has significant influence on the intrinsic properties of these crystals, including solubility and biocompatibility. This study aims to verify the possibility of functionalized hydroxyapatite nanoparticles and evaluate possible benefits of using these during the adhesive process in dentin. HA with different sizes and morphology were synthesized by precipitation method in aqueous solution. Furthermore, the influence of two new ligands, deoxycholic acid (AD) and abietic acid (AB) in the preparation of HA NP were investigated. The NP were characterized by analysis of EDX, DRX, DLS, TEM, MEV and AFM. The results showed that the addition of these ligands change the morphology but not the crystalline arrangement of HA. To study the interaction of NP of HA with collagen, NP of HA modified with AD was used. Bovine teeth were used to analyse the influence of using these NP during bonding procedures. The bovine teeth were divided into four groups: control (C), thermocycled Control (CTC), deoxycolic (AD) and deoxycolic thermocycled (ADTC) groups. The shear test was performed 24 hours after the adhesive procedure and after thermocycling (10,000 cycles). The adhesive interface images were performed before and after thermocycling by confocal Raman spectroscopy. The results showed an increase in initial resistance values of AD group when compared with group C, and no reduction in value after thermal cycling at ADTC group, which occurred in the CTC group. No change in the pattern of the hybrid layer was observed, showing a possible benefit of use this nanomaterial in dentistry.