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Avaliação do potencial remineralizador e propriedades mecânicas de um sistema adesivo experimental com análogos biomiméticos e fosfatos de cálcio bioativos : Assessment of remineralizing potential and mechanical properties of an experimental adhesive system mediated by biomimetic analogs and bioactive calcium phosphates / Assessment of remineralizing potential and mechanical properties of an experimental adhesive system mediated by biomimetic analogs and bioactive calcium phosphatesAbuna, Gabriel Flores, 1989- 27 August 2018 (has links)
Orientadores: Mário Alexandre Coelho Sinhoreti. Americo Bortolazzo Correr / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-27T04:06:35Z (GMT). No. of bitstreams: 1
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Previous issue date: 2015 / Resumo: Objetivos: O objetivo no presente trabalho foi avaliar o potencial remineralizador e a as propriedades adesivas (resistência de união, análise das interfaces, nanoinfiltração) de um novo sistema adesivo baseado em fosfatos e dois análogos biomiméticos incorporados em um adesivo experimental autocondicionante de dois passos. Métodos: Foi preparado um primer-base, onde foram incorporados os análogos e um adesivo-base contendo os fosfatos. Para avaliar o potencial remineralizador foram desmineralizadas barras de dentina (n=5) (EDTA 17%), distribuídas em: Controle sem análogos e sem fosfatos; Primer+Fosfatos; Acido poliacrilico APA+Fosfatos; Trimetafosfato de sódio TMS+Fosfatos; e Analogos+fosfatos. tratadas com cada primer e fixados junto a uma barra do adesivo polimerizado, em 1,5 mL de solução fosfato tampão. Foi usado um Espectroscópio Infravermelho Transformado de Fourier (ATR-FTIR) para caracterizar os fosfatos formados. Os espectros foram obtidos depois das 24 h, 7 dias, 2 e 6 meses. Microscopia Eletrônica de Transmissão (MET) foi usada para avaliar a interação dos cristais de apatita com o colágeno. Para avaliar as propriedades adesivas 50 dentes molares humanos hígidos foram usados (n=5): O primer foi aplicado por 10 segundos e o adesivo por 20 segundos, seguido de fotoativação por 20 s. Após, foram restaurados (Filtek Z350) em incrementos de 2 mm. Após 24 h ou 6 meses de pressão pulpar simulada (PPS), os dentes foram cortados em palitos de 0.9 x 0.9 mm2 e foi testada a resistência da união (EZ-test; Shimadzu, Kyoto, Japan), analisado o modo de fratura em estereomicroscópio. Um palito por dente foi selecionado e avaliado no ensaio nanoinfiltração, usando nitrato de prata amoniacal. Ainda, foi selecionado um palito por dente para avaliar a camada híbrida em Microscópio Eletrônico de Varredura com 15 KV. O analise estatístico da resistência de união foi feita com teste ANOVA dois fatores, e normalizados com Teste de Tukey, (p<0.05) Resultados: Todos os espetros do ATR-FTIR foram normalizados em 1650 cm-1 Amida I. Foi observado após 6 meses de PPS, a presença de picos em 947 e o par em 1092 cm-1 atribuídos a união C=O da HAp, e picos de PO4 em 1033 cm-1 , além de outros picos representativos da HAp. A análise em MET mostrou que, sem a intervenção dos análogos, a mineralização só ocorre nos espaços extrafibrilares, contrário a imagem com ambos análogos, onde observou-se uma organização em forma de agulhas intrafibrilar das apatitas. A resistência de união demostrou como a presença dos análogos e fosfatos não atrapalhou a adesão mostrando uma resistência similar no grupo de Analogos+Fosfatos, comparando-se 24 h com 6 meses de PPS (35,12±5,16 MPa e 38,67 ±4,03 MPa, respectivamente). No MEV observou-se a qualidade da camada adesiva, mostrando os túbulos dentinários obstruídos após 6 meses de PPS. Após a análise da nanoinfiltração, observou-se que a sorção de água diminui proporcionalmente à mineralização do grupo Analogos+Fosfatos. Conclusão: O sistema remineralizador proposto formou núcleos de HAp caracterizados mediante o ATR-FTIR, alcançando zonas intrafibrilares do colágeno, e mantendo o desempenho do adesivo após armazenamento em pressão pulpar simulada / Abstract: Objective: The aim of this study was to assess the remineralization potential and bonding properties of a novel system based on phosphates and biomimetic analogs incorporated into a two-step self-etch adhesive. Methods: It was prepared a main primer where were added the analogs and one main blend where were incorporated the phosphates. To assess the remineralization properties, to assess the mineral deposition dentin slabs were (n=5) demineralized (EDTA 17%), treated in each primer and fixed next to a polymerized adhesive slab (Under 1.5 mL phosphate buffer solution). It was used an ATR-FTIR to characterize the new phosphates. The spectra were recorded after 24 hrs, 7 days, 2 months and 6 months. The Transmission electronic microscopy was used to assess the nanometric interaction of apatite crystals and demineralized dentin collagen. In order to assess Dentin bonding properties, 50 non-carious human molars were used. (Under 038/2014 CEP-FOP protocol) divided into 5 groups n=10: PAA+Phosphates- polyacrylic acid-containing primer and phosphates-doped bond; STMP+Phosphates- Sodium trimetaphosphate-containing primer and phosphates-doped bond; Analogs+Phosphates- both analogs in the primer and phosphates-doped bond. The primer was applied for 10 s, the adhesive was applied for 20 s and light cured for 20 s. Then were resin (Filtek z350) restored at 2 mm layers. After 24 hrs or 6 months of simulated pulpal pressure SPP, the teeth were sectioned in sticks of 0.9 mm2 tested the µTensile Bond Strength, and analyzed the failure mode at a stereomicroscope. One stick per teeth was chosen and assessed Nanoleakage, using ammoniacal silver nitrate solution to reveal the hybrid layer at a Scanning Electronic Microscopy. Results: All the spectra of ATR-FTIR were normalized at 1650 Amide I. It was observed after 6 months of SPP, the presence of peaks at 947 and its pair at 1092 cm-1 attributed to C=O of HAp, and peaks of PO4 at 1033 cm-1 besides other specific peaks that confirm the presence of HAp. The TEM analysis showed, without the intervention of analogs the mineralization occurs just at in the extrafibrilar spaces, opposite to the image with both analogs where it is showed an intrafibrilar organization of needles like apatites. The µTBS outcomes demonstrate how the presence of this analogs and phosphates did not jeopardize the bonding strength. Showing a µTBS similar at Analogs+Phosphates, comparing 24 hrs of SPP to 6 months of SPP (35.12+5.16 MPa and 38.67 +4.03 MPa) (?=0.05). With SEM we observed the quality of this adhesive layer, showing occluded tubules after 6 months of SPP and no degradation of Resin tags. After Nanoleakage evaluation we see how after remineralization the water income decrease proportional to the mineralization. Conclusion: The proposed remineralizing system nucleate HAp characterized by ATR-FTIR in this study. Achieving the intrafibrilar zones of collagen, and keeping the performance of the adhesive after storage under SPP, maintaining the bonding properties and decreasing the factors of common adhesive degradation / Mestrado / Materiais Dentarios / Doutor em Materiais Dentários
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Chemical Controls on the Formation of Amorphous and Crystalline Calcium PhosphatesHoeher, Alexandria Janson 26 August 2020 (has links)
Transformation of amorphous calcium phosphate (ACP) and brushite into hydroxylapatite, an important biomineral, has been documented. The relationships between synthesis conditions and the formation and transformation of these phases are not comprehensively understood. The metastable nature of ACP has made it historically challenging to investigate, and many analyses attempt to stabilize the phase through drying or including additional ions or proteins in the reaction. In situ investigations provide an incisive approach to examining the structure and transformation of ACP and brushite as a function of synthesis conditions.
The first project develops a new method for in situ analyses of the structure of ACP and brushite shortly after reagent mixing, without chemical stabilization. This method was used in the second project to examine how the initial Ca/P affects ACP structure and transformation. Our results identify the first structural differences in types of ACP, controlled by the initial Ca/P. At ratio 0.2 the Ca – P bonding geometry is primarily monodentate, ratio 5.0 produces a coordination that is primarily bidentate, and there is a mix of monodentate and bidentate coordinates at intermediate ratios between the two. These results are independent of system pH between the examined range of 6-11. Further ex situ transformation experiments identified that at ratio 0.2, ACP transformed directly into hydroxylapatite, but at higher ratios the transformation product is brushite. This is a promising mechanism for direct ACP to hydroxylapatite conversion at a biologically relevant pH. In the final project, the statistically significant synthesis parameters (age, pH, temperature, supersaturation, and initial ion ratio) for ACP, brushite, and hydroxylapatite formation are evaluated. Binary logistic regression analysis and nonlinear continuous logistic regression analysis are applied to a dataset compiled from the literature. Equations were developed that predict the percentage of ACP and brushite that will form. The equations and significant variables seem to depend on the transformation pathway of brushite and ACP. The current analysis did not comprehensively describe hydroxylapatite formation when trying to concurrently evaluate the ACP to hydroxylapatite and brushite to hydroxylapatite pathways.
Taken together, these studies provide new ways to study and interpret calcium phosphate phases as they form and transform. Experiments identified new relationships between the chemistry and structure of ACP. The new in situ experimental method and the equations we developed can be used to improve future experimental designs towards a comprehensive understanding of the calcium phosphate system. / Doctor of Philosophy / Hydroxylapatite is a mineral made of calcium and phosphate, that is similar to the mineral components of bones and teeth in humans and other mammals and fish. Hydroxylapatite and other calcium phosphate phases can form when solutions, rich in calcium and phosphate, are mixed. Phases without long-range crystal structure, are amorphous calcium phosphates (ACP). Additional calcium phosphate minerals, like brushite can also form. If brushite and ACP are left in solution, they will transform into hydroxylapatite over time. Major questions include the need to learn the short-range atomic structure of ACP and how ACP and brushite transform into hydroxylapatite
In this dissertation, I investigate how chemistry and other variables such as age and time impact the calcium phosphate phase to form and how it transforms with aging. The first project develops a new method to study the structure of ACP and brushite without drying the study materials or adding additional chemicals or proteins to prevent them from transforming. The sample forms in a solution and flows directly through an X-ray beam for structural analysis.
This method is used in project two to examine how the ratio of calcium and phosphate in the beginning of the reaction affected the structure of ACP and how it transformed. The results identify the first structural differences in types of ACP, controlled by the initial Ca/P. At a ratio of 0.2 a calcium and phosphorus atom mostly share only one oxygen between them, but at Ca/P = 5.0, they mostly share two oxygens. At ratios in between 0.2 and 5.0 they share a mix of one and two oxygens. The results are independent of pH. Additionally, at ratio 0.2, ACP transformed directly into hydroxylapatite, but at all other ratios it transformed to brushite. Investigations of direct ACP to hydroxylapatite transformation are usually performed at a pH above that found in humans, but the transformation at low ratio occurred at a biologically relevant pH.
In the final project statistical analysis was used to identify what synthesis conditions (out of age, pH, temperature, supersaturation, and initial ion ratio) had a significant impact on the formation of ACP, brushite, and hydroxylapatite. Equations were developed that can be used to predict the percentage of ACP and brushite that form based on the statistically significant variables. The current analysis did not fully describe hydroxylapatite formation. Results suggest separate equations are needed for hydroxylapatite forming directly from ACP and directly from brushite.
Combined, these studies have created new ways to study calcium phosphate phases as they form and transform. This work experimentally identified new relationships between the chemistry and structure of ACP. Both the method and equations will help researchers improve their future experimental designs so investigations can be more directly compared to create a comprehensive understanding of calcium phosphates.
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POLYELECTROLYTE MULTILAYERS: SIMULATIONS, EXPERIMENTS, AND APPLICATIONS IN BIOMINERALIZATIONPatel, Pritesh A. January 2008 (has links)
No description available.
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Establishing a Baseline for Kinetic and Thermodynamic Origins of Vital Effects: Toward an Understanding of Factors Controlling Mg Signatures in CalciteStephenson, Allison Elaine 11 June 2009 (has links)
Elemental proxy models for temperature and seawater chemistry begin by assuming compositional signatures reflect environmental conditions of formation. The Mg/Ca ratio in marine cements and calcified skeletal structures is a widely used proxy for reconstructing past earth environments. Many studies have positively correlated Mg content in biogenic carbonates with temperature, but it is difficult to differentiate the effect of temperature from other environmental factors. Supersaturation, precipitation rate, salinity, pH, and ion concentration have also been proposed as drivers of Mg/Ca. Furthermore, it is difficult to distinguish environmental signatures from the “vital effect,” or the influences superimposed by the growth needs and metabolic activities of the organism. To construct viable paleoenvironmental proxies from biomineral compositions, we must resolve the effects of environmental conditions from the vital effects of the organism by first understanding the underlying thermodynamic and kinetic mechanisms for incorporating minor and trace elements.
Using in situ Atomic Force Microscopy, controlled solution chemistries, and different ion microprobe techniques, this dissertation investigates the kinetics and thermodynamics of calcite growth to establish an inorganic baseline for uptake of Mg. I use this information to quantify the enhancement in Mg/Ca due to the presence of hydrophilic 27-mer peptides, demonstrating a possible origin of vital effects. Likewise I measure the effect of ionic strength on signatures and find that growth rate and background electrolyte proved more important than salinity in determining Mg contents.
The findings contribute to the ongoing discussion regarding the relative importance of unique seawater parameters in determining Mg/Ca in calcite. Mg contents are significantly enhanced by biomolecules relative to the amounts attributed to temperature differences, while Mg content is less influenced by salinity variation than by changing supersaturation or driving force. In addition to sorting out the relative importance of environmental factors, our results begin to address the interplay of these different parameters in concert, and at different scales. At sites of calcification, the local biochemistry within an organism may shift in response to more saline waters. At a geological scale, interpreting past temperatures and particularly those of the Last Glacial Maximum depends on our ability to sort out and account for this interplay of salinity and temperature on Mg/Ca. Processes underlying inorganic and biogenic carbonate mineralization and interpretations of their formation environments are better understood by examining the influence of environmental parameters and biomolecular chemistry on kinetics and thermodynamics of calcite growth and stability. / Ph. D.
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Nanoscale Effects of Strontium on Calcite Growth: A Baseline for Understanding Biomineralization in the Absence of Vital EffectsWilson, Darren Scott 11 June 2003 (has links)
This study uses in situ atomic force microscopy (AFM) to directly observe the atomic scale effects of Sr on the monomolecular layer growth of abiotic calcite. These insights are coupled with quantitative measurements of the kinetics and thermodynamics of growth to determine the direction-specific effects of Sr on the positive and negative surface coordination environments that characterize calcite step edges.
Low concentrations of strontium enhance calcite growth rate through changes in kinetics. A new conceptual model is introduced to explain this behavior. Higher concentrations of strontium inhibit and ultimately stop calcite growth by a step blocking mechanism. The critical supersaturation required to initiate growth (sigma*) increases with increasing levels of strontium. At higher supersaturations, strontium causes growth rates to increase to levels greater than those for the pure system. The step blocking model proposed by Cabrera and Vermilyea in 1958 does not predict the experimental data reported in this study because the dependence of sigma* upon strontium concentration is not the same for all supersaturations.
Strontium inhibits calcite growth by different mechanisms for positive and negative step directions. Preliminary evidence indicates that strontium is preferentially incorporated into the positive step directions suggesting that impurity concentrations are not homogeneous throughout the crystal structure. Despite geochemical similarities, this study demonstrates that strontium and magnesium have different surface interaction mechanisms.
The findings of this study demonstrate the importance of understanding microscopic processes and the significance of interpreting biominerals trace element signatures in the context of direction-specific interactions. / Master of Science
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Establishing a physical and chemical framework for Amorphous Calcium Carbonate (ACC) biomineralizationMergelsberg, Sebastian Tobias 05 July 2018 (has links)
Recent advances in high-resolution analytical methods have brought about a paradigm shift in our understanding of how crystalline materials are formed. The scientific community now recognizes that many earth materials form by multiple pathways that involve metastable intermediates. Biogenic calcium carbonate minerals are now recognized to develop by aggregating molecules or clusters to form amorphous phases that later transform to one or more crystalline polymorphs. Amorphous calcium carbonate (ACC) is now recognized as a precursor to CaCO₃ biominerals in a wide variety of natural environments. Recent studies suggest an ACC pathway may imprint a different set of dependencies from those established for classical growth processes. Previous ACC studies provided important insights, but a quantitative understanding of controls on ACC composition when formed at near-physiological conditions is not established. The Mg content of ACC and calcite is of particular interest as a minor element that is frequently found in final crystalline products in calcified skeletons.
This three-part dissertation investigated biological and well-characterized synthetic ACC using high-energy x-ray methods, Raman spectroscopy, and mechanical tests. The findings establish chemical and physical properties of ACC in the exoskeleton of crustaceans and show Mg and P levels are tuned in the mineral component to optimize exoskeleton function that could be sensitive to ecological or environmental conditions. Calcite and chitin crystallinity exhibit a similar body-part-specific pattern that correlates directly with the mechanical strength of the exoskeleton. Insights from this study suggest precise biological control of ACC chemistry in the to regulate exoskeleton properties. Laboratory measurements using quantitative methods and compositions that approximate the physiological conditions of crustaceans, demonstrate at least two types of ACC are formed by controlling Mg concentration and alkalinity. We also find temporal changes in the short-range ordering of ACC after precipitation that is dependent upon carbonate content. The findings from this study provide a quantitative basis for deciphering relationships between ACC structures, solution chemistry, and the final transformation products under biologically relevant conditions. / Ph. D. / With the development of new imaging methods for nano-scale materials, scientists across diverse disciplines have recognized that many earth materials can form complex shapes by the formation and aggregation of nanocrystals or structureless (amorphous) particles. Biological minerals, such as shells and skeletons, are well-documented to form CaCO₃ via both of these attachment pathways, particularly amorphous calcium carbonate (ACC). However, little is known about the ACC properties and the factors that determine the final composition of skeletal minerals. This three-part dissertation focuses on ACC and calcite in the exoskeletons of crustaceans to understand how animals form composite exoskeletons of calcium carbonate minerals. This knowledge is important because CaCO₃ minerals are the primary component of the shells and skeletons of many economically important marine species. These minerals are also prevalent in the geological record as roadmaps for the evolutionary record. Amorphous and crystalline forms of CaCO₃ are also used as inert 'filler' materials for pharmaceutical products. By designing a series of experiments to characterize ACC in exoskeletons from lobsters and crabs, one part of the dissertation shows relationships between chemical composition and physical behavior of the materials. Building on this biomineral information, a separate experimental study synthesizes ACC under near-physiological conditions to show how amorphous CaCO₃ forms under controlled conditions. The findings have far-reaching consequences for understanding the complex chemistry that underlies the formation of calcium carbonate as a component of shells and skeletons, and what physical properties are optimized by the composition of these materials.
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Biomineralization of inorganic nanostructures using protein surfacesBergman, Kathryn N. 01 April 2008 (has links)
In nature, organisms have long been able to create elaborate mineral structures at ambient temperatures. From a materials science and engineering perspective, favorable properties emerge when the synthesis process can be controlled at finer levels. New strategies in materials chemistry synthesis has been inspired by biomineralization: biomimetics. In this work, silk fibroin films were used to synthesize gold nanoparticles room temperature by soaking a free standing 15nm silk film in HAuCl4. Particles ranged in size and shape from 5nm spheres to 105nm hexagons. Secondly, a film of ZnO1 peptide (ZnO selectively binding peptide) was successfully formed by drop casting on both silk and polystyrene surfaces. Using a HMT + Zn(NO3)2 system for ZnO wet chemical deposition, rods were formed on the peptide surface. Changing solution concentration and growth time affected the density and size of the nanorods. Spin coating a 3nm peptide film reduced the roughness to <1nm, upon which an array of vertical ZnO rods with controllable density was synthesized.
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Nonreductive biomineralization of uranium(VI) as a result of microbial phosphatase activityBeazley, Melanie J. 06 July 2009 (has links)
Uranium contamination of soils and groundwater at Department of Energy facilities across the United States is a primary environmental concern and the development of effective remediation strategies is a major challenge. Bioremediation, or the use of microbial enzymatic activity to facilitate the remediation of a contaminant, offers a promising in situ approach that may be less invasive than traditional methods, such as pump and treat or excavation.
This study demonstrates for the first time the successful biomineralization of uranium phosphate minerals as a result of microbial phosphatase activity at low pH in both aerobic and anaerobic conditions using pure cultures and soils from a contaminated waste site. Pure cultures of microorganisms isolated from soils of a low pH, high uranium- and nitrate-contaminated waste site, expressed constitutive phosphatase activity in response to an organophosphate addition in aerobic and anaerobic incubations. Sufficient phosphate was hydrolyzed to precipitate 73 to 95% total uranium as chernikovite identified by synchrotron X-ray absorption spectroscopy and X-ray diffraction. Highest rates of uranium precipitation and phosphatase activity were observed between pH 5.0 and 7.0.
Indigenous microorganisms were also stimulated by organophosphate amendment in soils from a contaminated waste site using flow-through reactors. High phosphate concentrations (0.5 to 3 mmol L-1) in pore water effluents were observed within days of organophosphate addition. Highest rates of phosphatase activity occurred at pH 5.5 in naturally low pH soils in the presence of high uranium and nitrate concentrations. The precipitation of uranium phosphate was identified by a combination of pore water measurements, solid phase extractions, synchrotron-based X-ray spectroscopy, and a reactive transport model.
The results of this study demonstrate that uranium is biomineralized to a highly insoluble uranyl phosphate mineral as a result of enzymatic hydrolysis of an organophosphate compound over a wide range of pH, in both aerobic and anaerobic conditions, and in the presence of high uranium and nitrate concentrations. The nonreductive biomineralization of U(VI) provides a promising new approach for in situ uranium bioremediation in low pH, high nitrate, and aerobic conditions that could be complementary to U(VI) bioreduction in high pH, low nitrate, and reducing environments.
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Calcium intake, physical activity, and bone mineral status in children and youth aged ten to fifteen yearsHall, Matthew Charles, 1960- January 1988 (has links)
This study was conducted to determine if either calcium intake or physical activity is related to bone mineral status in children and youth aged 10 to 15 years. Subjects (n = 30) with high, medium, and low bone mineral status were selected based on radius bone mineral index measurements from a sample of 108 subjects measured 9 to 12 months previously. Calcium intake was estimated from two 24-hour recalls and a food frequency questionnaire. Assessment of activity level was conducted by questionnaire. Single photon absorptiometry was used to obtain bone mineral content (g/cm) and bone mineral index (g/cm²) measurements for the radius and ulna at the midshaft and distal sites. Calcium intake and activity level were found to be similar among the bone mineral index groups. Using regression analysis, however, calcium intake was shown to be significantly related to midshaft ulna bone mineral index and activity level (sports participation) was found to be significantly related to distal ulna bone mineral index.
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Caractérisations physico-chimiques des biominéraux carbonatés de Mollusques actuels et fossiles : le cas des structures entrecroisées / Physico-chemical characterizations of carbonate biominerals from modern and fossil Molluscs : the case of intercrossed structuresNouet, Julius 09 April 2014 (has links)
Parmi la diversité des types microstructuraux que l’on peut trouver dans les coquilles des Mollusques, les architectures entrecroisées (lamellaire-croisé, lamellaire-croisé complexe, folié-croisé) sont de loin les plus abondantes. Mais elles restent, de par leurs organisations tridimensionnelles complexes, peu documentées : la majorité des études de biominéralisation, et les différents modèles qui en découlent, sont en effet essentiellement basés sur des architectures plus simples (telles que les couches nacrées ou prismatiques). Or il convient, si l'on souhaite aboutir à une meilleure compréhension des mécanismes fondamentaux qui dirigent la minéralisation de ces biocarbonates, de vérifier dans quelle mesure les modèles développés à partir de coquilles ”simples” sont aussi applicables à des organisations microstructurales plus complexes. Cette étude se focalise donc sur les couches entrecroisées des coquilles de quelques Mollusques, dans le but de mettre en évidence les différents niveaux du contrôle biologique que l'organisme exerce sur leur formation et leur croissance. À cette fin, des techniques de caractérisation in-situ des assemblages organiques sont privilégiées, en relation étroite avec l’analyse des organisations microstructurales et des mécanismes de biocristallisation à fine échelle. Quelques aspects de la diagénèse de ces microstructures seront aussi abordés, à travers l’étude de coquilles fossiles de Patella sp (~100 ka) et Velates perversus (~50 Ma). / Among the variety of microstructural types that can be found within Molluscs shells, intercrossed structures (crossed-lamellar, complex crossed-lamellar and cross-foliated) are by far the most commonly found. They are, however, still poorly documented - mainly due to their complex 3D organization. The majority of biomineralization studies, and the resulting models, are indeed essentially based on simple architectures (such as nacreous or prismatic layers). In order to achieve a better understanding of the fundamental mechanisms that drive the mineralization of such biocarbonates, it is therefore mandatory to check to which extent models developed from « simple » shells stay consistent when applied to more complex microstructural organizations. The present study focuses on intercrossed layers of several Mollusk shells, in order to highlight the various levels of biological control exerted by the organism on their formation and growth. In-situ techniques are used to characterize biochemical compositions, in close correlation with microstructural patterns, as well as fine-scale biocrystallization processes. Some peculiar features of the diagenesis of these microstructures are illustrated, through the study of fossil shells from Patella sp (~100 ky) and Velates perversus (~50 My).
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