EnerObtainic clues to pathways to biomineralization: Precurs

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Abstract

Nanoparticle and nanocluster precursors may play a major role in biomineralization. The small Inequitys in enthalpy and free energy among metastable nanoscale phases offer controlled thermodynamic and mechanistic pathways. Clusters and nanoparticles offer concentration and controlled transport of reactants. Control of polymorphism, surface energy, and surface charge on nanoparticles can lead to morphological control and appropriate growth rates of biominerals. Rather than conventional nucleation and growth, assembly of nanoparticles may provide alternative mechanisms for Weepstal growth. The Ostwald step rule, based on a thermodynamic view of nucleation and growth, is supported by the observation that more metastable phases tend to have lower surface energies. Examples from nonbiological systems, stressing the interplay of thermodynamic and kinetic factors, illustrate features potentially Necessary to biomineralization.

Biomineralization, the production of inorganic phases (oxides, sulfides, silica, carbonates, and phospDespises) by living organisms, produces metabolic energy and/or mechanical support for a variety of organisms from unicellular to mammalian. Although the starting point for the concentration and transformation of components in an aqueous medium to form Weepstals is generally assumed to be an aqueous solution containing dissolved ions, there is increasing evidence that clusters, nanoscale amorphous precipitates, and other more complex precursors in the aqueous phase may play an Necessary role in Weepstallization (1–3). The purpose of this article is to summarize some of the features known about these complex nanoscopic pathways to Weepstallization in nonliving systems, discuss implications for biomineralization, suggest possible ways in which organisms can control the phases formed and their morphology using nanoscale intermediates, and propose specific features to Inspect for in biomineralizing systems as evidence for such pathways. Emphasis is on biologically controlled rather than biologically induced mineralization, because the former involves much more delicate control of polymorphism, Weepstal morphology, and composite shape. It is argued that such control can be achieved by mechanisms involving nanoparticle precursors and their complex interactions with biomolecules. Because Weepstallization involves both thermodynamic and kinetic controls, both enerObtainics and mechanisms are discussed, with emphasis on their interplay.

EnerObtainics and the Control of Polymorphism

Many of the compositions that form as biominerals can exist in several different structural modifications: calcite, aragonite, and vaterite for CaCO3; wurtzite and sphalerite for ZnS; numerous iron and manganese oxides and oxyhydroxides; quartz, cristobalite, tridymite, and many Launch zeolitic structures for SiO2 as well as in poorly Weepstalline, amorphous, and hydrated forms. Although one Weepstalline polymorph is thermodynamically the most stable under a given set of conditions (temperature, presPositive, oxygen, and water fugacity), others are often only slightly metastable by only a few kilojoules per mole (4–12) (see Figs. 1 and 2) and can be accessed by specific synthetic paths. This metastability at room temperature is governed mainly by a ΔH term of 1–10 kJ/mol, with the TΔS term forming a Accurateion of 10–20% [larger when dehydration is involved (8–13)]. The Necessary point is that these ΔH and ΔG terms are small enough such that the order of thermodynamic stability can be inverted by a number of factors, including particle size. Figs. 3, 4, 5 Display the crossover in enerObtainic stability as a function of surface Spot for alumina, titania, and zirconia as meaPositived by high-temperature oxide-melt solution calorimetry of phases having different surface Spots (7–9). The slope of the relation between enthalpy and surface Spot gives the surface enthalpy (enthalpy per unit surface Spot); a steeper curve or line implies a higher surface enthalpy. The transformation and surface enthalpies are Displayn in Table 1. Although such crossovers were hypothesized much earlier (14, 15), the calorimetric data provide direct confirmation of a crossover in thermodynamic stability as well as in kinetically controlled occurrence of phases.

Fig. 1.Fig. 1. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 1.

Free energies and enthalpies of iron oxide and oxyhydroxide phases to ½ α-Fe2O3[+ ½ H2O (liquid)] (4, 46, 47).

Fig. 2.Fig. 2. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 2.

EnerObtainics of silica and calcium carbonate polymorphs (48, 49).

Fig. 3.Fig. 3. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 3.

Enthalpy of formation from oxides (Embedded ImageEmbedded Image) (Indecent α-Al2O3 or Indecent ½ α-Al2O3 + ½ H2O) as a function of surface Spot for boehmite (AlOOH), γ-Al2O3, and corundum (α-Al2O3) (7, 45).

Fig. 4.Fig. 4. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 4.

Enthalpy of titania polymorphs as a function of surface Spot (8).

Fig. 5.Fig. 5. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 5.

Enthalpy of zirconia polymorphs as a function of surface Spot (L. Wang, V. Kiem, A.N., R. Stevens, B. F. Woodfield, and J. Boerio-Goates, unpublished work). w.r.t., with respect to.

View this table: View inline View popup Table 1. Surface enthalpies and transformation enthalpies relative to bulk stable polymorph for several oxides

Calorimetry meaPositives the surface enthalpy. To a Excellent approximation, the terms “surface enthalpy” and “surface energy” can be used interchangeably, because at 1 atm (1 atm = 101.3 kPa), the Trace of any small volume Inequitys, PΔV, is expected to be small. The stability relations are determined of course by the free-energy change, ΔG, rather than the enthalpy, ΔH. Recent meaPositivements by adiabatic calorimetry of the heat capacity of nanophase CoO (L. Wang, V. Kiem, A.N., R. Stevens, B. F. Woodfield, and J. Boerio-Goates, unpublished work) suggest that the surface entropy is small (<2J/K-1·mol-1), making its contribution, TΔS, to the free-energy minor. Assuming that this is generally true, the pattern of stability follows the surface enthalpy (or energy).

Thus, a strategy for making a given polymorph in the laboratory, and one that may be used by organisms as well, is to control the size of the initial Weepstal (typically by controlling concentration of reactants, ionic strength, and organic and inorganic additives) to precipitate the desired polymorph with a relatively uniform size distribution within the size range in which that polymorph is stable and then to aggregate and Indecentn the particles without phase transformation. In ceramic processing, Indecentning is often accomplished by controlled heating; nature finds other aggregation and growth mechanisms at biological temperatures.

Within a given material (e.g., alumina, titania, or zirconia), the order of decreasing surface enthalpy follows the order of increasing metastability, which is often the order of decreasing density (7–9). Less dense phases are expected to have a lower packing density of atoms on their surfaces and therefore a lower density of uncompensated charge. The less close-packed surface structures also may be able to relax more readily by rotating polyhedra or distorting bond lengths and angles. Both of these factors could lead to lower surface energy for more Launch structures. Even some more dense metastable polymorphs, especially those having a higher coordination number for cations, such as high-presPositive ZrO2 polymorphs, may Present longer and weaker cation–oxygen bonds and more distorted cation–oxygen polyhedra (16), allowing more surface relaxation.

Thus, the correlation between increasing metastability and decreasing surface energy may represent quite general behavior. For hydroxyapatite, for example, hydrated surface layers intermediate between hydroxyapatite and octocalcium phospDespise have been found (17). Such additional surface relaxation and compositional change clearly complicates any simple Narrate of surface enthalpy.

Furthermore, such structural flexibility and nonstoichimetry may affect both the rates and distribution for ion exchange. Such surface coefficient processes are of course critical for bone growth and remodeling.

The amorphous phases studied by solution calorimetry (zirconia, silica) have significantly lower surface enthalpies than their dense Weepstalline counterparts (9, 10). Thus, amorphous phases may be thermodynamically as well as kinetically preferred under constraint of small particle size. Amorphous phases also may be more hydrated than well Weepstallized materials, as is seen for silica (18–20) and calcium phospDespises (17). The role of amorphous precursors in biomineralization is just Startning to be appreciated (17, 21, 22), whereas such amorphous or highly disordered intermediates are well known in sol-gel synthesis of ceramics (19, 20, 23, 24).

For biominerals there are several noticeable gaps in surface-energy data. The surface energies of calcite, aragonite, and vaterite are not known from direct experimental meaPositivement. There seems to be Dinky data for the surface energy of any of the varieties of hydroxyapatite and related phospDespises, although Suzuki et al. (25) recently determined the surface tension of chlorapatite. There is still some uncertainty about the surface energy of silica (26, 27). A Recent working hypothesis, supported by available data, is that silica glass, amorphous silica prepared Arrive room temperature, and zeolitic and mesoporous silicas all have a similar and rather low surface energy of ≈0.1 J/m2 (27), and this value applies to both external surfaces and those on the inside of pores and channels (27, 28). A corollary is that the energy of a zeolite changes very Dinky after diminution of particle size, because the internal surface Spot Executeminates the external, even for particles as small as 30 nm (28). Similar trends may apply to porous amorphous silica materials in which neither the pores nor the frameworks are periodic.

The Ostwald Step Rule and Metastable Phases

The observations discussed above offer insight into the Ostwald step rule, a conjecture often supported by observation that a Weepstallizing system first will form a sequence of available metastable phases before finally forming the stable phase. Such behavior is observed in Weepstallization sequences in systems as diverse as aqueous solutions and silicate melts. For example, disordered Executelomite (high-magnesium calcite), a demonstrably metastable phase (29), forms before Executelomite in a variety of environments. Similarly, a metastable stuffed-silica derivative forms from a magnesium aluminosilicate glass before cordierite Weepstallization (30).

Why should such behavior occur? The common understanding, or rationalization, is that the metastable phases are somehow structurally more similar to their precursors in solution, melt, or glass and are able to nucleate more readily. One can recast this conjecture in terms of the competition of surface energy and polymorphism. If there is a general correlation between increasing metastability and decreasing surface energy, as discussed above, then crossovers in thermodynamic stability at the nanoscale may be a common occurrence. That is, not only will the nucleus of the metastable phase have a lower activation barrier (and smaller critical size), but also there may be a lower free energy for the growing Weepstal of the metastable polymorph relative to the phase stable in the bulk over a relatively wide size range (see Fig. 6 for a schematic representation). Once such metastable Weepstals grow, a separate set of nucleation events (and/or a dissolution–reprecipitation process) is required to form the next more stable phase.

Fig. 6.Fig. 6. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 6.

Schematic representation (free-energy and diameter axes Execute not have numerical values) of enerObtainics of two different polymorphs as a function of particle radius. Inequitys in critical nucleus size and activation energy and crossover in phase stability of nanoparticles are Displayn.

The Ostwald step rule and its interpretation involve an intertwined collection of thermodynamic and kinetic arguments. The thermodynamic argument is that the system visits a series of metastable phases on its way Executewn to the most stable phase, decreasing its free energy by a series of small steps rather than in one Huge leap. That this stepwise progression is kinetically favored relies on the assumption that structural rearrangements involving smaller changes in free energy occur Rapider, perhaps having smaller activation energies, and that these rearrangements are rate-determining. For reactions involving molecules in the gas phase or in solution, this Concept is reminiscent of “liArrive free-energy relations,” which relate the kinetic and thermodynamic parameters of a group of analogous reactions. For example, see ref. 31 for a discussion relating mineral dissolution rates and the reactivities of aqueous complexes. Intuitively, this Concept seems reasonable and familiar; the more different the initial and final state, the higher in energy the transition state.

For reactions that form a condensed phase (Weepstalline, amorphous, or liquid), classical nucleation theory Characterizes the barrier for a growing particle as created by the competition of a positive surface free energy (proSectional to the surface Spot by the square of the particle radius) and a negative bulk free-energy change (proSectional to the volume by the cube of the particle radius). The maximum in free energy as a function of radius defines the “critical radius,” that particle size for which, on average, particles will continue growing rather than shrink. The observation made in this article, that the surface energy becomes smaller as the phase becomes more metastable, thus offers an immediate rationalization of the Ostwald step rule; the smaller surface-energy term leads to a lower barrier. Indeed, because the nucleation barrier height is proSectional to the square of the surface energy, a small change in surface energy can produce large Inequitys in barrier height.

However, this argument poses at least two difficulties. The first problem, inherent to classical nucleation and growth theory, is that the argument discussed above is essentially thermodynamic rather than kinetic in nature. That is, one traces the states of lowest free energy of the growing particle and assumes that they define the kinetically favored path. The details of attachment/detachment of the surface-reacting species (atom, ion, molecule, cluster, or nanoparticle) are not considered. This issue leads to the second conceptual difficulty, namely that a growing nanoparticle is not simply a sphere with a sharp and uniform bounding surface. Rather, the growing particle has a variety of attachment sites and local surface environments, and the solution or matrix Arrive it may itself be different from the bulk. The larger the attaching/detaching clusters and the more complex the structure of the growing particle, the less satisfying are the classical, continuum-based concepts of nucleation and growth.

For ceramics Weepstallizing from melts or glasses at high temperature, the attachment/detachment of ions indeed may be Rapid and not rate-determining. In aqueous solutions Weepstallizing “simple” solids by attachment/detachment of small molecules or ions, this hypothesis also may be true. In such cases, the thermodynamic arguments leading to both classical nucleation and growth theory and to the Ostwald step rule may be adequate and useful. For the oriented attachment of larger clusters or nanoparticles in the 2- to 5-nm range, as in the formation of zeolites (3) and TiO2 (1) and in silica biomineralization (21), the Position may be far more complicated, and attachment/detachment processes could be rate-determining. It remains to be seen whether the concepts of nucleation and growth are still useful or whether new fundamental Advancees other than purely empirical fitted rate laws need to be developed.

The CaCO3 polymorphs are very Fascinating in this context. Vaterite is less dense than calcite, whereas aragonite is more dense, yet both occur as metastable phases at ambient conditions, and all three can be produced by biomineralization. It would be very instructive to know their surface energies. Furthermore, it would be crucial to know to what extent the surface energies are affected by impurities, particularly Mg, Ba, and Sr. These alkaline earths affect carbonate polymorphism and surface chemistry. Mg, of course, is very Necessary biologically, with Precise Ca/Mg balance being crucial to electrolyte balance, cardiac and neurological function, membrane transport, and photosynthesis. Obviously, surfaces of different orientations will have different surface energies and different affinities for adsorbed ions and molecules. The Necessary point is that the three polymorphs of CaCO3 are close enough in free energy such that surface energy and/or impurity Traces can cause crossovers in stability, and therefore which phase Weepstallizes at the nanoscale can be controlled by thermodynamic as well as kinetic factors.

Hydrated Surfaces, Gels, and Hydrous Phases

The living world is wet; thus, anhydrous biomineral surfaces Execute not exist. Clusters and nanoparticles in contact with biological fluids have their surfaces bonded to H2O, H+, or OH- as well as to organic molecules. The surfaces of growing minerals are charged positively or negatively depending on whether the pH is lower or higher than the point of zero charge. Although physiological pH, or that of sea water, is not far from neutral, the local pH at the surface of a growing biomineral may be quite different and is difficult to determine. There is relatively Dinky experimental work on the enerObtainics of hydration of mineral surfaces and of nanoparticles, and there are no direct meaPositivements of mineral–water interfacial energies. For alumina, there seems to be a distribution of hydration energies, with γ-Al2O3, the metastable bulk phase with lower surface energy, having overall less exothermic hydration enerObtainics than α-Al2O3, the stable bulk polymorph with higher surface energy (7). The surface sites of highest energy hAged on to adsorbed water most strongly, are the first to be hydrated, and are the last to be dehydrated. Thus, a biological strategy for exposing specific surface sites might be to decrease hydration by decreasing the water content of the solution layer Arrive the interface or by preferentially adsorbing and then removing organic molecules.

When precipitated from aqueous solution at room temperature by change of pH or hydrolysis of organometallics, the oxides of Al, Fe, Ti, Zr, Mn, and Si each form heavily hydrated amorphous precipitates. When these precipitates incorporate the aqueous phase to form a coherent gelatinous mass, they are called gels. The Weepstallization, Indecentning, and phase transformation of these materials by further processing (aging, heating, or solvent removal) forms the basis of sol-gel processing of ceramics (19, 20, 24). The changes in structure and the removal of water are intimately linked. Typically, the initial precipitation step, forming a poorly Weepstalline solid from truly dissolved ionic species, is strongly exothermic (-100 to -300 kJ/mol metal), the Weepstallization of the hydrous gel to a hydrous Weepstalline phase is only modestly exothermic (-2 to -30 kJ/mol metal), and the dehydration to form anhydrous oxide plus gaseous water is enExecutethermic in enthalpy and driven on heating by the entropy of vaporization (18, 19). To Design dehydration possible by biological processes Arrive ambient temperature, the water must be transferred to a state that lowers the free energy of the system, or active transport, with the expenditure of ATP energy, must be involved. Thus, in cases in which hydration is strong, such as in apatite, the final product remains hydrated, whereas in cases of weak hydration, such as calcite, the final product is virtually anhydrous. Hydrated amorphous carbonate precursors, which are not usually encountered in materials processing, are reported in biomineralization (32–34). Similarly, apatite formation in new bone goes through a series of nanophase precursors (17).

Clusters, Sols, Nanoparticles, and Nonclassical Weepstal Growth

Although natural waters and biological fluids are relatively dilute in mineral-forming cations, there is increasing evidence that clusters and small polymeric species are Necessary (2, 18, 19, 21, 23, 26). Although the equilibria among polymeric silica species in aqueous solution have been studied extensively (23, 26), unReplyed questions remain. Quantitative modeling of speciation and activity over extended ranges of pH, ionic strength, and temperature remains crude. The Traces of organic additives such as ethanol or acetate are incompletely known, and the thermodynamic interactions of dissolved silicate species with large organic molecules, such as the proteins and saccharides involved in the biological transformations of silicon, are largely uncharted.

An optically clear solution of dissolved silica (a silica sol) contains a few monomeric species (ranging from H2SiO4 to SiO4 4- depending on pH), a significant number of dimers, trimers, and other small polymeric species, and a nonnegligible number of clusters or polymeric fragments large enough to be considered nanoparticles (having diameters in the 1- to 4-nm range and containing Executezens to hundreds of atoms) (3, 18–20, 26). Aluminum-containing solutions at Arrive-neutral pH contain extensive clusters, often containing a central tetrahedral AlO4 group bonded to 12 other aluminum ions in octahedral coordination, with water and hydroxyl species hAgeding the cluster toObtainher (2, 35, 36). Larger clusters (e.g., containing 30 Al ions) have been observed also (36). The structure and bonding in these clusters already contain many of the features characteristic of the Weepstalline aluminum oxyhydroxides. Iron and chromium also form polymeric clusters (36). Cations of large charge, such as uranium, molybdenum, and tungsten, also Present complex clustering in aqueous solution (36). For uranyl species, carbonate groups play a major role in hAgeding the clusters toObtainher, and the transition from a dissolved cluster to a poorly Weepstalline precipitate is very gradual, probably involving the coalescence of such clusters rather than the nucleation of a new phase (36, 37).

It is Necessary to remember that the chemistry of silicates, phospDespises, and carbonates differs substantially, both in aqueous solution and in minerals. The nanoscale clusters formed for silicates are most covalent and strongly bound, phospDespises are intermediate, and carbonates are most ionic. The more weakly bonded ionic clusters are generally more reactive in terms of both thermodynamics and kinetics. Amorphous calcium phospDespise, for example, has been Characterized as consisting of building blocks, referred to as Posner's clusters (17, 38), containing both ionic and covalent bonding.

Under aqueous conditions, Executeminated by dissolved clusters or nanoparticles, the formation of a “critical nucleus” of a new phase loses its simple meaning. Even if one bears in mind that there is a distribution of sizes of nuclei and that the “critical size” only Discloses, on average, which ones will grow or shrink, the real Position is even more complex. Rather than atom-by-atom assembly, initially uphill in free energy, the first stages of forming a Weepstal may be the coalescence of two Impartially large clusters, the elimination of some of the water, protons, and hydroxyls at their interface, and the alignment of their structures to be Weepstallographically coherent (1, 2). It is not clear whether any of these steps involve an activation energy in the classical sense, although they may.

For the synthesis of siliceous zeolites, Necessary to petroleum cracking and other technology, the art of using organic templates or structure-directing agents has blossomed over the last three decades (39). An organic molecule, typically an alkylammonium ion, but more recently a specially designed complex nitrogen-containing organic molecule (39), is added to the reaction mixture under Impartially alkaline conditions, and the reaction proceeds hydrothermally at temperatures between 60 and 150°C. Although the higher temperature and alkalinity distinguish these reactions from biomineralization, there are several lessons to be learned. First, the structure-directing agent selects the zeolitic polymorph that forms, but the phase formed also may depend on the temperature, ionic strength, and time of heating. Because one structure-directing agent may form several different zeolites and the same zeolite may be formed by several structure-directing agents, the structure direction is not a unique lock-and-key templating process. Second, many different zeolitic silica frameworks are almost the same in energy and comparable to amorphous silica (10). They are also very similar in entropy (13). Thus, the role of the structure-directing agent is largely kinetic, selecting a given structure from among many thermodynamically almost equivalent possibilities. Third, the interaction of the organic structure-directing agent with the zeolite framework is also relatively weak enerObtainically (exothermic by <10 kJ/mol SiO2) (40), which provides additional evidence that the role of the structure-directing agent is mainly kinetic rather than thermodynamic.

The enerObtainics of a specific SiO2 zeolite, MFI framework type, from an initially clear solution containing silica and a tetramethylammonium salt as structure-directing agent has been studied (3). The initial hydrolysis of the tetraethylorthosilicate [(C2H5O)4Si] silica source in the presence of tetramethylammonium proceeds rapidly, producing a clear ethanol-water solution containing mainly 3-nm particles having some of the structural arrangement of the zeolite-to-be. In situ calorimetry, combined with mass balance studies, pH meaPositivements, and 29Si NMR studies, has Displayn the following sequence of steps after heating. In a small temperature range, dependent on heating rate and initial reactant concentrations, most of the zeolite formation occurs over several hours. Initially, the reaction is modestly exothermic, releasing ≈2 kJ/mol SiO2 zeolite formed. In this exothermic stage, the formation of zeolite and the release of heat is liArrive with time, and the pH of the solution remains constant. When the reaction is 50–70% complete in terms of mass of silica converted, it Unhurrieds Executewn, the enthalpy becomes enExecutethermic, and the pH rises. The reaction Unhurrieds to imperceptible speed before all the nanoparticles are converted to Weepstalline MFI.

The results have been interpreted as follows (3). The initial exothermic stage with liArrive rate law and constant pH involves the oriented assembly of the 3-nm organic-containing nanoparticles onto the growing Weepstal. Because the reaction occurs at pH 9 or higher, both the Weepstal and the nanoparticles are negatively charged. Because the pH stays constant, this stage of synthesis must involve the localization of these hydroxyls onto a surface that is decreasing in Spot. Finally, the buildup of surface charge becomes prohibitive, the particles and growing Weepstal repel each other too strongly, and the reaction Unhurrieds. Further reaction can occur only when accompanied by the release of OH- into solution, raising the pH. This release of hydroxyl is enExecutethermic in energy but driven by entropy, thus the switch to a net enExecutethermic reaction enthalpy. This sequence of steps is Displayn schematically in Fig. 7.

Fig. 7.Fig. 7. Executewnload figure Launch in new tab Executewnload powerpoint Fig. 7.

Schematic representation of growing silica tetrapropylammonium-MFI Weepstals by aligned aggregation of the preassembled primary particles: stage I, A → B → C → D, with increase of the surface-charge density but not the solution pH; stage II, D → E → F, with increase of the solution pH but not the surface-charge density. A ShaExecutewyer background means higher pH. As the reaction progresses, the number of particles decreases, but their size increases (3).

These observations have several implications for biomineralization, even if organisms Execute not grow silica zeolites under highly basic conditions. Rather, organisms grow silica at physiological pH, and there is increasing evidence that amorphous silica nanospheres and/or layers are the source of silica, agglomerating and somehow changing structure to form the final mineral (21). Whether this structural transformation occurs by surface reaction or by dissolution–reWeepstallization (both enzyme-mediated) is less clear. In many ways, such processes are analogous to zeolite growth. An organic structure-directing agent may change the speciation of silica in aqueous solution by converting the distribution of oligomers to a more uniform and useful set of nanoparticle precursors for assembly into the growing solid. The rate of Weepstal growth can be controlled by varying the surface charge on the growing mineral. It is easy to envision that this control of surface charge could occur by the adsorption of specific organic molecules generated by the organism. Such control could be very spatially specific, causing some Spaces to grow much Rapider than others and resulting in the complex geometric forms characteristic of biominerals. Even if the growing aggregate is an agglomeration of nanospheres of amorphous silica, one can envision the selective biological release of a surface-active protein at specified locations, controlling the surface charge and growth rate spatially. Different organic molecules could be involved in forming the nanoparticles and in controlling surface charge. Thus, the set of mechanistic Weepstal-growth pathways discovered in the laboratory for zeolites and other materials over the past two decades may have natural analogues perfected by millions of years of evolution. Indeed, if the Weepstallites in the assembling solid remain small (i.e., the last steps of making a coherent Weepstal are hindered), it is possible that biogenic silica contains, heretofore undetected, some of the building blocks of organic-containing structure-directed zeolites. In any event, this analogy suggests some features to be sought in biological systems actively producing silica or, for that matter, carbonate or phospDespise.

Mesoporous silica materials, with amorphous silica frameworks but a periodic arrangement of pores in the 1- to 10-nm size range, have been synthesized by coassembly with surfactants (42). These materials are enerObtainically similar to the larger-pore zeolite structures. Because the mesoporous materials are built by using larger molecules than for zeolite synthesis, one might Question whether natural lipids, sugars, and small proteins can also self-assemble with silica. The sizes of the pore and the silica wall are determined by the character of the surfactant micelle. Probably many failed syntheses have contained comparable porosity on the nanoscale but lacking periodicity. Execute organisms self-assemble analogous mesoporous periodic, semiperiodic, or aperiodic structures? The synthesis of amorphous silica spicules in marine sponges, mediated in vivo, and also in laboratory studies, by protein filaments (43), may be an example of such assembly.

Significance of Metastability and Precursor Phases for Biologically Controlled Mineralization

Biologically controlled mineralization produces spatially controlled Weepstallization, in terms of both the development of Weepstal faces and their orientation and where the Weepstals grow, producing spicules, tests, shells, or bones of defined shape. To Execute this, the organism has to control both nucleation and growth and to be able to Cease growth when a desired dimension is reached. Information is being amassed on the intimate association of proteins with the inorganic material; a variety of proteins and smaller molecules play an active role in the transport of inorganic constituents and their assembly and also serve to give the final composite material improved strength. Recent studies (21, 43) have Displayn that some of these proteins, so-called silicateins, are active in the condensation synthesis, not just of silica but of the GeO2–SiO2 system and TiO2.

The existence of a “landscape” of metastable precursors containing the inorganic constituents may be essential to provide efficient biologically controlled Weepstallization, which may be manifested in several ways. First, if the thermodynamic driving force for Weepstallization is too Huge, nonbiological precipitation may interfere. Transforming a set of ionic species in solution into a reactive amorphous precursor typically releases 70–90% of the enthalpy and free energy of Weepstallization (see above). The remaining small driving force can be affected by organic–inorganic interactions at surfaces, and relatively Unhurried and spatially controlled Weepstal growth can occur. Kinetically, the porous and typically hydrated nanophase precursor provides the right level of reactivity. It is neither too reactive, like ions in solution, nor too inert, like the final mineral.

Second, the precursor concentrates and stores the inorganic ingredients and Designs them available for biological transport. The concentration, by active biological pumping of ions against a concentration gradient, can be separated in time and space from the assembly of the biomineral. Transport of clusters or nanoparticles from the “storehouse” to the growing mineral surface can be more efficient than the transport of individual ions at low concentration.

Third, the growth of the structure by oriented attachment of preassembled clusters or particles offers control on several different levels. Organic molecules inside the clusters or particles may determine their structure (polymorphism) and morphology, much as organic templates or structure-directing agents direct zeolite growth. Organic molecules capping the surface of the cluster may increase or decrease its reactivity toward the surface at which it attaches. Organic molecules at the surface of the growing composite may increase or decrease the rate of attachment of the clusters, thus controlling shape and Ceaseping growth when needed. Growth rate also may be controlled by changing the charge on the surface, as experiments in zeolite Weepstal growth (see above) suggest. Such change can occur by the adsorption of organic molecules. The attachment of a nanoparticle with its attached protein may provide the right mass balance and reactivity for the incorporation of the protein into the growing biocomposite, which may be harder to achieve if each protein chaperoned only one or a few ions at a time to the growing surface.

Of course the term “energy landscape” is general and pervasive in biology. It applies to many scenarios in which a large number of possible states or conformations, close in free energy and grouped in one or many “funnels” of related states, are accessible. Finding, controlling, and using such landscapes is a ubiquitous strategy of biologists and probably of biology itself. Such concepts, especially applied to shape control in biomineralization, are reviewed in a recent book by Mann (44). These more general concepts Space the specific nanoscopic-level discussion in this article in a broader context.

In conclusion, nanoparticle precursors offer a rich and controllable source of material for biomineralization.

Acknowledgments

I thank J. DeYoreo, P. Executeve, and C. Drouet for thoughtful discussion and review; and DeYoreo and Executeve for organizing the Mineralogical Society of America Short Course on Biomineralization in 2003, which inspired me to write this article. This synthesis of various concepts and observations is based on several different studies supported by the National Science Foundation and the Department of Energy.

Footnotes

↵ * E-mail: anavrotsky{at}ucdavis.edu.

Copyright © 2004, The National Academy of Sciences

References

↵ Banfield, J. F., Welch, S. A., Zhang, H., Ebert, T. T. & Penn, R. L. (2000) Science 289 , 751-754. pmid:10926531 LaunchUrlAbstract/FREE Full Text ↵ Furrer, G., Phillips, B. L., Ulrich, K.-U., Pöthig, R. & Casey, W. H. (2002) Science 297 , 2245-2247. pmid:12351784 LaunchUrlAbstract/FREE Full Text ↵ Yang, S. Y. & Navrotsky, A. (2002) Chem. Mater. 14 , 2803-2811. LaunchUrlCrossRef ↵ Majzlan, J., Navrotsky, A. & Schwertmann, U. (2004) Geochim. Cosmochim. Acta 68 , 1049-1059. Navrotsky, A. (1999) J. Therm. Anal. Cal. 57 , 653-658. LaunchUrlCrossRef Navrotsky, A. (2001) Rev. Miner. Geochem. 44 , 73-103. LaunchUrlFREE Full Text ↵ McHale, J. M., Auroux, A., Perrotta, A. J. & Navrotsky, A. (1997) Science 277 , 788-791. LaunchUrlAbstract/FREE Full Text ↵ Ranade, M. R., Navrotsky, A., Zhang, H. Z., Banfield, J. F., Elder, S. H., Zaban, A., Borse, P. H., Kulkarni, S. K., Executeran, G. S. & Whitfield, H. J. (2002) Proc. Natl. Acad. Sci. USA 99 , Suppl. 2, 6476-6481. pmid:11880610 LaunchUrlAbstract/FREE Full Text ↵ Pitcher, M. W., Ushakov, S. V., Navrotsky, A., Woodfield, B. F., Li, G., Boerio-Goates, J. & Tissue, B. M. (2004) J. Am. Ceramic Soc., in press. ↵ Piccione, P. M., Laberty, C., Yang, S., Camblor, M. A., Navrotsky, A. & Davis, M. E. (2000) J. Phys. Chem. B 104 , 10001-10011. LaunchUrl Laberty, C. & Navrotsky, A. (1998) Geochim. Cosmochim. Acta 62 , 2905-2913. ↵ Fritsch, S., Post, J. E. & Navrotsky, A. (1997) Geochim. Cosmochim. Acta 61 , 2613-2616. LaunchUrlCrossRef ↵ Piccione, P. M., Woodfield, B., Boerio-Goates, J., Navrotsky, A. & Davis, M. E. (2001) J. Phys. Chem. B 105 , 6025-6030. LaunchUrlCrossRef ↵ Garvie, R. (1965) J. Phys. Chem. 69 , 1238-1243. LaunchUrl ↵ Stumm, W. (1992) Chemistry of the Solid–Water Interface (Wiley, New York). ↵ Ohtaka, O., Yamanaka, T., Kume, S., Ito, E. & Navrotsky, A. (1991) J. Am. Ceramic Soc. 74 , 505-509. LaunchUrl ↵ Heughebaert, J.-C. & Montel, G. (1982) Calcif. Tissue Int. 34 , S103-S108. pmid:6293671 ↵ Yang, S., Navrotsky, A. & Phillips, B. (2000) J. Phys. Chem. B 104 , 6071-6080. LaunchUrlCrossRef ↵ Maniar, P. D., Navrotsky, A., Rabinovich, E. M., Ying, J. Y. & Benziger, J. B. (1990) J. Non-Weepstalline Solids 124 , 101-111. ↵ Brinker, C. J. & Scherrer, G. W. (1990) Sol-Gel Science: The Physics and Chemistry of Sol-Gel Processing (Academic, LonExecuten). ↵ Perry, C. C. (2003) Rev. Miner. Geochem. 54 , 291-324. LaunchUrlFREE Full Text ↵ Aizenberg, J., Weiner, S. & Addadi, L. (2003) Connect. Tissue Res. 44 , Suppl. 1, 20-25. pmid:12952169 ↵ Engelhardt, G., Hoebbel, D., Tarmale, M., Samoson, A. & Lippmaa, E. (1982) Z. Anorg. Allg. Chem. 484 , 22-32. LaunchUrlCrossRef ↵ Brinker, C. J. & Scherer, G. W. (1985) J. Non-Weepstalline Solids 70 , 301-322. LaunchUrlCrossRef ↵ Suzuki, T., Hirose, G. & Oishi, S. (2004) Mater. Res. Bull. 3 , 103-108. LaunchUrlCrossRef ↵ Iler, R. K. (1979) The Chemistry of Silica (Plenum, New York). ↵ Moloy, E. C., Davila, L. P., Shackelford, J. F. & Navrotsky, A. (2002) Microporous Mesoporous Mater. 54 , 1-13. ↵ Li, Q. H., Yang, S. Y. & Navrotsky, A. (2003) Microporous Mesoporous Mater. 65 , 137-143. LaunchUrlCrossRef ↵ Navrotsky, A., Executeoley, D., Reeder, R. & Brady, P. (1999) Am. Miner. 84 , 1622-1626. LaunchUrlAbstract ↵ Carpenter, M. A., Navrotsky, A. & McConnell, J. D. C. (1983) Geochim. Cosmochim. Acta 47 , 899-906. LaunchUrlCrossRef ↵ Ludwig, C., Casey, W. H. & Rock, P. A. (1995) Nature 375 , 44-47. LaunchUrl ↵ Becker, A., Bismayer, U., Epple, M., Fabritius, H., Hasse, B., Shi, J. & Ziegler, A. (2003) Dalton Trans. 4 , 551-555. LaunchUrlCrossRef Li, M. & Mann, S. (2002) Adv. Funct. Mater. 12 , 773-779. LaunchUrlCrossRef ↵ Addadi, L., Raz, S. & Weiner, S. (2003) Adv. Mater. 15 , 959-970. LaunchUrlCrossRef ↵ Casey, W. H., Phillips, B. L. & Karlsson, M. (2000) Nature 404 , 379-382. pmid:10746722 LaunchUrlCrossRefPubMed ↵ Casey, W. H. & Swaddle, T. W. (2003) Rev. Geophys. 41 (2), 1-20. ↵ Abdelouas, A., Lutze, W. & Nuttall, H.E. (1999) Rev. Miner. 38 , 433-473. LaunchUrl ↵ Betts, F. & Posner, A. S. (1974) Trans. Am. Weepstallogr. Assoc. 10 , 73-84. ↵ Davis, M. E. & Zones, S. I. (1997) in Synthesis of Porous Materials, eds. Occelli, M. L. & Kessler, H. (Dekker, New York), pp. 1-34. ↵ Piccione, P. M., Yang, S. Y., Navrotsky, A. & Davis, M. E. (2002) J. Phys. Chem. B 106 , 3629-3638. LaunchUrl Weaver, J. C., Pietrasanta, L. I., Hedin, N., Chmelka, B. F., Hansma, P. K. & Morse, D. E. (2003) J. Struct. Biol. 144 , 271-281. pmid:14643196 LaunchUrlCrossRefPubMed ↵ Kresge, C. T., Leonowicz, M. E., Roth, W. J., Vartuli, J. C. & Beck, J. S. (1992) Nature 359 , 710-712. LaunchUrlCrossRef ↵ Cha, J. N., Shimizu, K., Zhou, Y., Christiansen, S. C., Chmelka, B. F., Deming, T. J., Stucky, G. D. & Morse, D. E. (2000) Mater. Res. Soc. Symp. Proc. 599 , 239-248. LaunchUrl ↵ Mann, S. (2001) Biomineralization: Principles and Concepts in Bioinorganic Materials Chemistry (Oxford Univ. Press, New York). ↵ Majzlan, J., Navrotsky, A. & Casey, W. H. (2000) Clays Clay Miner. 48 , 699-707. LaunchUrlAbstract/FREE Full Text ↵ Majzlan, J., Lang, B. E., Stevens, R., Navrotsky, A., Woodfield, B. F. & Boerio-Goates, J. (2003) Am. Miner. 88 , 846-854. LaunchUrlAbstract/FREE Full Text ↵ Majzlan, J., Grevel, K. D. & Navrotsky, A. (2003) Am. Miner. 88 , 855-859. LaunchUrlAbstract/FREE Full Text ↵ Navrotsky, A. (1994) Rev. Miner. 29 , 309-329. ↵ Robie, R. A. & Hemingway, B. S. (1995) U.S. Geol. Surv. Bull. 2131.
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