Crystalline order of a water/glycine film coadsorbed on the (104) calcite surface

For biomineralization processes, the interaction of the surface of calcite crystals with organic molecules is of particular importance. Especially, biologically controlled biomineralization as in exoskeletons of mollusks and echinoderms, e.g., sea urchin with single-crystal-like spines and shells,1-3 requires molecular control of seed formation and growth process. So far, experiments showing the obvious influence of organic molecules on the morphology and habit of calcite crystals have demonstrated the molecular dimension of the interaction.4-7 Details of the kinetics of growth and dissolution of mineral surfaces influenced by additives are available,8,9 but other experimental data about the structure of the organic/inorganic interface on the atomic scale are rare. On the other hand, complicated organic macromolecules which are involved in biomineralization are numerous, with only a small fraction solved in structure and function so far.10-13 Therefore, model systems have to be designed to provide a basic understanding for the interaction process.14 Using grazing incidence X-ray diffraction combined with molecular modeling techniques, we show that glycine molecules order periodically on the calcite (104) face in competition with the solvent water when exposed to an aqueous solution of the most simple amino acid. In contrast to the general concept of the charge-matching fit of organic molecules on mineral surfaces,4,14 glycine is not attached to the calcite surface directly but substitutes for water molecules in the second hydration layer.     

草酸钙型尿结石病人结石和尿液微晶的形貌特征分析

泌尿系结石是临床上的常见病和多发病,可生长于肾盂、肾实质、输尿管以及膀胱中,较大的结石常会导致尿道阻塞、感染甚至肾脏坏死等病变,是一种严重危害人体健康的泌尿系统疾病。尿结石是动物体内异常生物矿化的产物,其主要成分有草酸钙、磷酸铵镁、尿酸等,其中以草酸钙最为常见。本研究直接取肾结石病人排出体外的自然结石和尿液中的结石微晶作为研究对象,采用透射电子显微镜、扫描电子显微镜、X-射线衍射分析和傅立叶变换红外光谱等手段对尿结石进行表征分析。研究结果表明,尿液中的纳米微晶的聚集是导致微晶快速增大和结石形成的关键因素,通过调控纳米微晶的物理化学性质有可能抑制肾结石的形成和复发。     

A promising structural zinc enzyme model for CO2 fixation and calcification

A new ligand, namely, N-{tris([2-[(1-methylbenzimidazol-2-yl)ethyl]methyl]amino)-2-oxoethyl}iminodiacetic acid is synthesized and characterized and used to prepare a zinc complex as a promising model for the active site of the nacreous protein in mollusc shells. Preliminary investigation of CO₂ fixation and calcification is studied with regard to the influence of the pK_a value of the coordinated water molecule and the carboxylate groups.     

Acidic Monosaccharides become Incorporated into Calcite Single Crystals**

Carbohydrates, along with proteins and peptides, are known to represent a major class of biomacromolecules involved in calcium carbonate biomineralization. However, in spite of multiple physical and biochemical characterizations, the explicit role of saccharide macromolecules (long chains of carbohydrate molecules) in mineral deposition is not yet understood. In this study, we investigated the influence of two common acidic monosaccharides (MSs), the two simplest forms of acidic carbohydrates, namely glucuronic and galacturonic acids, on the formation of calcite crystals in vitro. We show here that the size, morphology, and microstructure of calcite crystals are altered when they are grown in the presence of these MSs. More importantly, these MSs were found to become incorporated into the calcite crystalline lattice and induce anisotropic lattice distortions, a phenomenon widely studied for other biomolecules related to CaCO₃ biomineralization, but never before reported in the case of single MSs. Changes in the calcite lattice induced by MSs incorporation were precisely determined by high-resolution synchrotron powder X-ray diffraction. We believe that the results of this research may deepen our understanding of the interaction of saccharide polymers with an inorganic host and shed light on the implications of carbohydrates for biomineralization processes.     

From Bacteria to Mollusks: The Principles Underlying the Biomineralization of Iron Oxide Materials

Various organisms possess a genetic program that enables the controlled formation of a mineral, a process termed biomineralization. The variety of biological material architectures is mind‐boggling and arises from the ability of organisms to exert control over crystal nucleation and growth. The structure and composition of biominerals equip biomineralizing organisms with properties and functionalities that abiotically formed materials, made of the same mineral, usually lack. Therefore, elucidating the mechanisms underlying biomineralization and morphogenesis is of interdisciplinary interest to extract design principles that will enable the biomimetic formation of functional materials with similar capabilities. Herein, we summarize what is known about iron oxides formed by bacteria and mollusks for their magnetic and mechanical properties. We describe the chemical and biological machineries that are involved in controlling mineral precipitation and organization and show how these organisms are able to form highly complex structures under physiological conditions.     

Lattice Shrinkage by Incorporation of Recombinant Starmaker-Like Protein within Bioinspired Calcium Carbonate Crystals

The biological mediation of mineral formation (biomineralization) is realized through diverse organic macromolecules that guide this process in a spatial and temporal manner. Although the role of these molecules in biomineralization is being gradually revealed, the molecular basis of their regulatory function is still poorly understood. In this study, the incorporation and distribution of the model intrinsically disordered starmaker-like (Stm-l) protein, which is active in fish otoliths biomineralization, within calcium carbonate crystals, is revealed. Stm-l promotes crystal nucleation and anisotropic tailoring of crystal morphology. Intracrystalline incorporation of Stm-l protein unexpectedly results in shrinkage (and not expansion, as commonly described in biomineral and bioinspired crystals) of the crystal lattice volume, which is described herein, for the first time, for bioinspired mineralization. A ring pattern was observed in crystals grown for 48 h; this was composed of a protein-enriched region flanked by protein-depleted regions. It can be explained as a result of the Ostwald-like ripening process and intrinsic properties of Stm-l, and bears some analogy to the daily growth layers of the otolith.     

Mineral formation on dentin induced by nano-bioactive glass

The object of this study was to evaluate the effect of bioactive glass (BG) size on mineral formation on dentin surfaces. Totally demineralized dentin discs were treated using BG suspensions with different particle sizes:i.e., microscale bioactive glass (m-BG), submicroscale bioactive glass (sm-BG) and nanoscale bioactive glass (n-BG). Field-emission scanning electron microscopy and 3D profile measurement laser microscopy were used to observe the surface morphology and roughness. It was found that all BG particles could promoted mineral formation on dentin surfaces, while plug-like depositions were observed on the dentin discs treated by n-BG and they were more acid-resistant. The present results may imply that n-BG has potential clinical application for dentin hypersensitivity treatment.     

Biomimetic Mineralization of Protein Nanogels for Enzyme Protection

Protein nanogels have found a wide variety of applications, ranging from biocatalysis to drug/protein delivery. However, in practical applications, proteins in nanogels may suffer from enzymic hydrolysis and denaturation. Inspired by the structure and functionalities of the fowl eggshells, biomimetic mineralization of protein nanogels was studied in this research. Protein nanogels with embedded porcine pancreas lipase (PPL) in the cross-linked nanostructures were synthesized through the thiol–disulfide reaction between thiol-functionalized PPL and poly(N-isopropylacrylamide) with pendant pyridyl disulfide groups. The nanogels were further reacted with reduced bovine serum albumin (BSA) and BSA molecules were coated on the nanogels. Mineralization of BSA leads to the synthesis of biomineralized shells on the nanogels. With the growth of CaCO₃ on the shells, the nanogels aggregate into suprastructures. Thermogravimetric analysis, XRD, dynamic light scattering, and TEM were employed to study the mechanism of the biomineralization process and analyze the structures of the mineralized nanogels. The biomineralized shells can effectively protect the PPL molecules from hydrolysis by trypsin; meanwhile, the nanosized channels on the mineralized shells allow the transport of small-molecule substrates across the shells. Bioactivity measurements indicate that PPL in the nanogels maintains more than 80 % bioactivity after biomineralization.     

Ab Initio structure determination of vaterite by automated electron diffraction

"This is a mineral about which there has been much discussion" is a typical statement about vaterite in older standard textbooks of inorganic chemistry. This polymorph of CaCO(3) was first mentioned by H. Vater in 1897, plays key roles in weathering and biomineralization processes, but occurs only in the form of nanosized crystals, unsuitable for structure determination. Its structure could now be solved by automated electron diffraction tomography from 50 nm sized nanocrystals.     

Bioinspired growth of crystalline carbonate apatite on biodegradable polymer substrata

Mineralization in biological systems is a widespread, yet incompletely understood phenomenon involving complex interactions at the biomacromolecule-mineral nucleus interface. This study was aimed at understanding and controlling mineral formation in a poly(alpha-hydroxy ester) model system, to gain insight into biological mineralization processes and to develop biomaterials for orthopaedic tissue regeneration. We specifically hypothesized that providing a high surface density of anionic functional groups would enhance nucleation and growth of bonelike mineral following exposure to simulated body fluids (SBF). Polymer surface functionalization was achieved via hydrolysis of 85:15 poly(lactide-co-glycolide) (PLG) films. This treatment led to an increase in surface carboxylic acid and hydroxyl groups, resulting in a substantial increase in polymer surface energy from 42 to 49 dynes/cm2. Treated polymers exhibited a 3-fold increase in heterogeneous mineral grown and growth of a continuous mineral film on the polymer surface. The mineral grown on PLG surfaces is a carbonate apatite, the major mineral component of vertebrate bone tissue. Mineral crystal size and morphology were dependent on the solution characteristics but unaffected by the degree of surface prehydrolysis. The mechanism of heterogeneous carbonate apatite growth was examined via ion binding assays, which indicated that calcium binding is mediated independently by the presence of soluble phosphate counterions and surface functional groups. These findings indicate that poly(alpha-hydroxy ester) materials can be readily mineralized using a biomimetic process, and that the impetus for mineral nucleation in this system appears more complicated than the simple electrostatic interactions proposed in previous biomineralization theory.     

Structure, self-assembly, and dual role of a beta-defensin-like peptide from the Chinese soft-shelled turtle eggshell matrix

Biomineral matrix formation and molecular recognition are two important processes associated with eggshell biomineralization. To understand these two processes, a major intracrystalline peptide, pelovaterin, was isolated from turtle (Pelodiscus sinensis) eggshell and its tertiary and quaternary structures were established. The global fold of pelovaterin is similar to that of human beta-defensins but has a large hydrophobic core and a short hydrophilic N-terminal segment, which is not preserved in defensins. Pelovaterin exhibits strong antimicrobial activity against two pathogenic gram-negative bacteria, Pseudomonas aeruginosa and Proteus vulgaris, and stabilizes a thin film of metastable vaterite. We show that pelovaterin self-aggregates in the form of micellar nanospheres and the aggregation in solution is entropy-driven. It is suggested that the micellar aggregation of pelovaterin is responsible for the induction and stabilization of the metastable phase by altering the interfacial energy. The results demonstrate the adaptability of an extracellular matrix protein to perform multiple tasks: polymorph discrimination and protection of the contents of the egg against bacterial invasion.     

Dentin Phosphoprotein Mimetic Peptide Nanofibers Promote Biomineralization

Dentin phosphoprotein (DPP) is a major component of the dentin matrix playing crucial role in hydroxyapatite deposition during bone mineralization, making it a prime candidate for the design of novel materials for bone and tooth regeneration. The bioactivity of DPP‐derived proteins is controlled by the phosphorylation and dephosphorylation of the serine residues. Here an enzyme‐responsive peptide nanofiber system inducing biomineralization is demonstrated. It closely emulates the structural and functional properties of DPP and facilitates apatite‐like mineral deposition. The DPP‐mimetic peptide molecules self‐assemble through dephosphorylation by alkaline phosphatase (ALP), an enzyme participating in tooth and bone matrix mineralization. Nanofiber network formation is also induced through addition of calcium ions. The gelation process following nanofiber formation produces a mineralized extracellular matrix like material, where scaffold properties and phosphate groups promote mineralization. It is demonstrated that the DPP‐mimetic peptide nanofiber networks can be used for apatite‐like mineral deposition for bone regeneration.     

Spectroscopic characterization of mineral crystals produced by the bacterium Azospirillum brasilense

The results of spectroscopic structural and trace elemental analyses of mineral crystals produced by the soil nitrogen-fixing bacterium Azospirillum brasilense cultivated in a synthetic medium are presented and discussed. The mineral formed is shown to have a structure close to struvite (MgNH(4)PO(4) x 6H(2)O; ASTM file No. 15-762) with some differences which may be attributed to the presence of isomorphic admixtures of other cations (struvite is known to have a variety of forms). AAS/AES and ion chromatography analyses for a number of biologically important microelements and their role in the formation of the crystal structure, as well as some questions related to biomineralization are also discussed.     

Nanoscale Confinement Controls the Crystallization of Calcium Phosphate: Relevance to Bone Formation

A key feature of biomineralization processes is that they take place within confined volumes, in which the local environment can have significant effects on mineral formation. Herein, we investigate the influence of confinement on the formation mechanism and structure of calcium phosphate (CaP). This is of particular relevance to the formation of dentine and bone, structures of which are based on highly mineralized collagen fibrils. CaP was precipitated within 25–300 nm diameter, cylindrical pores of track etched and anodised alumina membranes under physiological conditions, in which this system enables systematic study of the effects of the pore size in the absence of a structural match between the matrix and the growing crystals. Our results show that the main products were polycrystalline hydroxapatite (HAP) rods, together with some single crystal octacalcium phosphate (OCP) rods. Notably, we demonstrate that these were generated though an intermediate amorphous calcium phosphate (ACP) phase, and that ACP is significantly stabilised in confinement. This effect may have significance to the mineralization of bone, which can occur through a transient ACP phase. We also show that orientation of the HAP comparable, or even superior to that seen in bone can be achieved through confinement effects alone. Although this simple experimental system cannot be considered, a direct mimic of the in vivo formation of ultrathin HAP platelets within collagen fibrils, our results show that the effects of physical confinement should not be neglected when considering the mechanisms of formation of structures, such as bones and teeth.     

Structures,organo–mineral compositions and diagenetic changes in biominerals

Numerous investigations on mineral–organic relationships in biominerals have recently been undertaken. They provide information on the biomineralization and diagenetic processes, and new insights on the important issue of the role of the organic matrices. Progress has come, in part, from improvements in analysis tools and from the combination of multi-technique observations.     

SEM–FTIR spectroscopic evaluation of deterioration in an historic coffered ceiling

Microorganisms generally degrade wood when moisture, oxygen and other environmental factors provide favorable growing conditions. Scanning electron microscopy (SEM) and Fourier transform infrared spectroscopy (FTIR) were used to study the development and transformation of the products formed during the biomineralization process that follows the deterioration of wood from an historical coffered ceiling (Cloister of St Francesc (XIV century), Palma de Mallorca, Spain). After fungi colonization, cellulose and lignin disappear and inorganic salts are formed. Thus, the secretion of numerous acids (initially oxalic acid) by fungal hyphae also leads to the precipitation of authigenic salts. Damaged cells or tissues enhance the formation and growth of crystals, which is strongly favored by fungi that function as calcification nuclei. Finally, the presence of dihydrated calcium sulfate reveals the contribution of environmental factors to the weathering of wood.     

The microstructures of biomineralized surfaces: a spectroscopic study on the exoskeletons of fresh water (Apple) snail, Pila globosa

In view of the importance in understanding biomineralization processes in different molluskan species, the common fresh water apple snail Pila globosa in Indian origin was taken to explore its mineralized exoskeleton structures. The detailed structural studies of the exoskeletons of P. globosa have been undertaken. The isolated layers present in these shells were studied by electron paramagnetic resonance (EPR), optical absorption, and infrared spectral techniques. The EPR spectra of the organic protein layer periostracum show the characteristic signals corresponding to Fe(3+) ions at g = 4.1 and 2.0. The EPR spectra of the ostracum (middle) layer at room temperature gives a complicated spectrum consisting of a number of Mn(2+) signals of at least three sets due to the aragonite nature of the material. The results indicate the presence of the multivalent manganese ions, which undergo the redox mechanisms. The thermal variation of the EPR spectra show marked effect on these samples both in g-values and the basic spectral pattern.     

高效液相色谱法制备人胆汁中结合胆红素

针对单结合胆红素（ｍｏｎｏｃｏｎｊｕｇａｔｅｄｂｉｌｉｒｕｂｉｎ，ＭＣＢ）理化性质极不稳定而造成对其分析、制备的困难性，在Ｓｐｉｖａｋ和Ｔｒｏｔｍａｎ方法的基础上建立了一个简单可行的人胆汁结合胆红素（ｃｏｎｊｕｇａｔｅｄｂｉｌｉｒｕｂｉｎ，ＣＢ）高效液相色谱（ＨＰＬＣ）二次洗脱的制备方法，并对方法的可靠性作了论证，得到了ＭＣＢ较为满意的量与纯度，为进一步探讨ＭＣＢ的理化性质及其与胆石形成的关系提供了条件。     

仿生矿化的机理和应用研究进展

碳酸钙、磷酸钙为代表的生物矿物广泛分布于自然界中,经过不同的矿化过程,在生物体内呈现出多样的结构、形貌和功能,构成生物体多种组织和器官.在人工材料合成领域,仿生矿化通过调控碳酸钙、磷酸钙等矿物的成核与生长,获得具有复杂高级结构和特殊生物功能的无机或无机/有机复合材料.本文重点介绍仿生矿化机理和应用的最近研究进展,包括仿...     

Application of scanning electron microscopy and X-ray microanalysis: FE-SEM, ESEM-EDS, and EDS mapping for studying the characteristics of topographical microstructure and elemental mapping of human cardiac calcified deposition

To explore the pathogenic mineral formation in a huge cardiolith isolated from the left heart atrium of an 80-year-old male patient, field emission scanning electron microscopy (FE-SEM) was used to analyze the topographic microstructure and perform elemental mapping in a cross-section of the cardiac calcified deposit after dissection. Environmental SEM equipped with an energy dispersive X-ray spectrometer (EDS) was also used to investigate the composition and spatial distribution of elements in the cross-section, and fiberoptic Raman spectroscopy was used to reidentify the chemical composition of designated positions. The results indicated that calcium hydroxyapatite and cholesterol were the main components of the cardiac calculus. The plate-like structures of calcium hydroxyapatite were unevenly spread over the cholesterol of the cardiac calculus. The calcium hydroxyapatite-rich area exhibited higher amounts of C, O, P, and Ca elements as well as trace amounts of N, Na, Mg, and Al, whereas the major concentration of C, minor concentrations of N and O, and trace amounts of P and Ca were observed in the cholesterol-rich area. Hypercholesterolemia associated with calcification of this cardiac calculus was proposed. Both FE-SEM and ESEM energy dispersive X-ray microanalyses were performed directly, for the first time, to provide useful information on the microstructural characteristics and spatial distribution of elements on the surface of human cardiac calculi.