Biomineral Crystals Research Articles

Asprich Peptides Are Occluded in Calcite and Permanently Disorder Biomineral Crystals

Macromolecules are a minority but important component of the minerals formed by living organisms, or biominerals. The role these macromolecules play at the early stages of biomineral formation, as well as their long-term and long-range effects on the mature biomineral, is poorly understood. A 42-amino acid peptide, asp2, was derived from the Asprich family of proteins. In this study we present X-ray absorption near-edge structure spectroscopy and X-ray photoelectron emission microscopy data from the asp2 peptide, the calcite (CaCO(3)) crystals, and the peptide + crystal composites. The results clearly show that asp2 is occluded in fully formed biomineral crystals and slightly but permanently disorders the crystal structure at short- and long-range distances.

Effects of crystalline phase on the biological properties of collagen–hydroxyapatite composites

The objective of this study was to investigate the effects of spatial structure and crystalline phase on the biological performance of collagen–hydroxyapatite (Col–HA) composite prepared by biomineralization crystallization. Two types of Col–HA composites were prepared using mineralization crystallization (MC composites) and pre-crystallization (PC composites), respectively. Structural characteristics were analyzed by scanning electron microscopy and transmission electron microscopy. Surface elemental compositions were measured by electron spectroscopy for chemical analysis (ESCA). These composites were used in in vivo repair of bone defects. The effects of the crystalline phase on the biological performance of Col–HA composites were investigated using radionuclide bone scan, histopathology and morphological observation. It was observed that in MC composites, HA was located on the surface of the collagen fibers and aggregated into crystal balls, whereas HA in PC composites was scattered among the collagen fibers. ESCA showed that phosphorus and calcium were 8.99% and 17.56% on MC composite surface, compared with 4.39% and 5.86% on the PC composite surface. In vivo bone defect repair experiments revealed that radionuclide uptake was significantly higher in the area implanted with the PC composite than in the contralateral area implanted with the MC composite. Throughout the whole repair process, the PC composite proved to be superior to the MC composite with regard to capillary-forming capacity and the amount of newly formed bone tissue. So it could be concluded that HA placement on collagen fibers affected the biological performance of Col–HA composites. Pre-crystallization made HA scattered among collagen fibers, creating a better structure for bone defect repair in comparison with MC Col–HA composites.

Photonic crystals based on natural oceanic biominerals

The photonic-crystal properties of natural biomineral crystals—siliceous basal spicules of glass sea sponges—have been studied for the first time. It is shown that the periodic arrangement of concentric cylindrical layers composed of nanodimensional silica particles leads to the formation of forbidden photonic bands.

Annealing-induced interfacial toughening using a molecular nanolayer

Self-assembled molecular nanolayers (MNLs) composed of short organic chains and terminated with desired functional groups are attractive for modifying surface properties for a variety of applications. For example, organosilane MNLs are used as lubricants, in nanolithography, for corrosion protection and in the crystallization of biominerals. Recent work has explored uses of MNLs at thin-film interfaces, both as active components in molecular devices, and as passive layers, inhibiting interfacial diffusion, promoting adhesion and toughening brittle nanoporous structures. The relatively low stability of MNLs on surfaces at temperatures above 350-400 degrees C (refs 12, 13), as a result of desorption or degradation, limits the use of surface MNLs in high-temperature applications. Here we harness MNLs at thin-film interfaces at temperatures higher than the MNL desorption temperature to fortify copper-dielectric interfaces relevant to wiring in micro- and nano-electronic devices. Annealing Cu/MNL/SiO2 structures at 400-700 degrees C results in interfaces that are five times tougher than pristine Cu/SiO2 structures, yielding values exceeding approximately 20 J m(-2). Previously, similarly high toughness values have only been obtained using micrometre-thick interfacial layers. Electron spectroscopy of fracture surfaces and density functional theory modelling of molecular stretching and fracture show that toughening arises from thermally activated interfacial siloxane bridging that enables the MNL to be strongly linked to both the adjacent layers at the interface, and suppresses MNL desorption. We anticipate that our findings will open up opportunities for molecular-level tailoring of a variety of interfacial properties, at processing temperatures higher than previously envisaged, for applications where microlayers are not a viable option-such as in nanodevices or in thermally resistant molecular-inorganic hybrid devices.

Surface structure study of biological calcium phosphate apatite crystals from human tooth enamel.

A surface-structure study of human tooth enamel crystals has been carried out by high-resolution electron microscopy (HREM). The surfaces of several crystals have been examined and the surfaces of a single crystal are described here. The observations made on this crystal are similar to the observations made on the other crystals, although the difference of morphology in the crystals observed indicates that the growth control by matrix proteins takes only place on the [011 macro 0] surfaces of the crystals. The crystal described here is oriented along the [112 macro 0] direction, and the following surfaces have been analyzed: (11 macro 00), (011 macro 1), (01 macro 11) and (0001). A subsurface reconstruction is observed just below the surface of the crystal not bonded to the matrix and lying above the supporting film. Observation of the matrix surrounding the crystal shows the existence of poorly crystalline phases with a structure close to that of hydroxyapatite (OHAP). Finally, a comparison of the images of the (011 macro 0) surface with computer-simulated images calculated for several models of the OHAP surface structure shows that the surface itself is stoichiometric and contains both calcium and phosphate groups. The knowledge of the binding sites of proteins on biomineral crystals such as the ones found in human tooth enamel is of prime importance for the understanding of their growth process. In this study, the first structure determination of the surface of human tooth enamel crystals is presented.

Incorporation of fluorescent molecules and proteins into calcium oxalate monohydrate single crystals

A central question to understanding biomineralization is determining how biomolecules are integrated within inorganic host lattices, thereby changing material properties yet retaining single crystal structure of the biomineral. We have addressed guest incorporation within single biomineral crystals by investigating face specificity, anisotropy and the role of charges in fluorescent calcium oxalate monohydrate (COM) crystals nucleated from solutions containing eosin Y, fluoresceins and rhodamines. Additionally, we have examined the specificity of incorporation of Protein G wild-type (G-wt) and its mutant (G-Δ6, in which four aspartic acids and two glutamic acids have been replaced by the corresponding asparagine or glutamine), where G-wt promotes and G-Δ6 inhibits COM crystal growth. We found that (1) the negatively charged fluorophores, as well as the fluorophore-labeled proteins, G-wt and G-Δ6, were successfully incorporated during growth into the same {1 1 0} growth sectors in preference to all others; (2) the positively charged TRITC (tetramethyl rhodamine isothiocyanate) was not incorporated as free fluorophore, but it became incorporated if conjugated to G-wt and G-Δ6; (3) once the fluorophores are incorporated, the polarization measurements of adsorption and emission were similar irrespectively whether taken from COM containing the free fluorophores, or the protein conjugates. The anisotropy was similar for rhodamines and fluoresceins.

Template induced crystallization of biominerals on self-assembled monolayers of alkylthiols

We have studied the templated crystallization of the three polymorphs of calcium carbonate on self-assembled monolayers of ω-substituted alkylthiols on gold surfaces. Our interest has been to model biomineralization processes by examining the effect of structured organic templates on heterogeneous nucleation at the inorganic–organic interphase. The crystallization of the three polymorphs was carried out from solution at two different temperatures. The crystals that were formed on the surface were examined using scanning electron microscopy. Phase quantification was achieved through Rietveld analysis of the X-ray diffraction profiles. This study provides significant clues on factors that affect such templated crystallization.