Dental composites based on hybrid and surface-modified amorphous calcium phosphates.

The objectives of this study were to prepare hybrid and surface-modified amorphous calcium phosphates (ACPs) as fillers for mineral-releasing dental composites, and determine whether the mechanical strength of the composites could be improved without decreasing their remineralization potential. ACP was hybridized with tetraethoxysilane or zirconyl chloride and surface-treated with 3-methacryloxypropoxytrimethoxy silane (MPTMS) or zirconyl dimethacrylate (ZrDMA). Composites fabricated with unmodified ACP (u-ACP), hybrid or surface-modified ACP filler and photo-activated Bis-GMA, TEGDMA and 2-hydroxyethyl methacrylate (HEMA) (BTH resin), Bis-GMA, TEGDMA, HEMA and MPTMS (BTHS resin) or Bis-GMA, TEGDMA, HEMA and ZrDMA (BTHZ resin) were tested for their remineralizing potential and biaxial flexure strength (BFS). Ion releases from all composites were significantly above the minimum necessary for reprecipitation of apatite. The BFS of unfilled polymers was not adversely affected by immersion in saline solutions. The BFS of BTH and BTHS composites deteriorated upon soaking. However, BTHZ composites were practically unaffected by exposure to saline solutions. Filler hybridization resulted in a modest, but significant, improvement in the BFS (up to 24%) of BTHZ composites. Heterogeneous distribution of the ACP on disk surfaces was detected by the FTIR microspectroscopy analyses. This might have been caused by uncontrolled aggregation of ACP particles that appeared to hinder interfacial filler/resin interactions and diminish the mechanical strength of composites.

Amorphous Calcium Phosphate-Based Bioactive Polymeric Composites for Mineralized Tissue Regeneration.

Amorphous calcium phosphate (ACP), a postulated precursor in the formation of biological hydroxyapatite, has been evaluated as a filler phase in bioactive polymeric composites that utilize dental monomers to form the matrix phase on polymerization. In addition to excellent biocompatibility, these composites provided sustained release of calcium and phosphate ions into simulated saliva milieus. In an effort to enhance the physicochemical and mechanical properties and extend the utility of remineralizing ACP composites to a greater variety of dental applications, we have focused on: a) hybridizing ACP by introducing silica and/or zirconia, b) assessing the efficacy of potential coupling agents, c) investigating the effects of chemical structure and compositional variation of the resin matrices on the mechanical strength and ion-releasing properties of the composites, and d) improving the intrinsic adhesiveness of composites by using bifunctional monomers with an affinity for tooth structure in resin formulations. Si- and Zr-modified ACPs along with several monomer systems are found useful in formulating composites with improved mechanical and remineralizing properties. Structure-property studies have proven helpful in advancing our understanding of the remineralizing behavior of these bioactive composites. It is expected that this knowledge base will direct future research and lead to clinically valuable products, especially therapeutic materials appropriate for the healing or even regeneration of defective teeth and bone structures.

Silica- and zirconia-hybridized amorphous calcium phosphate: effect on transformation to hydroxyapatite.

The goal of this study was to determine the effect that silica and zirconia have on the stability of bioactive amorphous calcium phosphate (ACP) mineral, i.e., in retarding its transformation to hydroxyapatite (HAP). The glass-forming agents, tetraethoxysilane and zirconyl chloride, were introduced individually during the low-temperature preparation of ACP. These hybrid ACPs (Si-ACP and Zr-ACP, respectively) as well as the control, unhybridized ACP (u-ACP), were characterized by X-ray diffraction, Fourier transform infrared spectroscopy, scanning electron microscopy, specific surface area measurements, and chemical analysis (Ca/PO(4) ratio of the solids) before being dispersed in one of the following four test solutions: N-2-hydroxyethylpiperazine-N'-2-ethanesulfonic acid (HEPES)-buffered (pH = 7.40) saline solutions with 0 microg/g fluoride (test solution A1), 1 microg/g fluoride (test solution A2), and 10 microg/g fluoride (test solution A3), or a lactic acid-containing solution (pH = 5.10, adjusted with NaOH; test solution B). Aliquots were taken at predetermined time intervals for solution Ca and PO(4) analysis. Solids isolated after 30 and 90 min exposure to solution B as well as the final dissolution/transformation products from all four solution experiments were analyzed by Fourier transform infrared spectroscopy and X-ray diffraction. Regardless of the type of experimental solution used, slower conversion to HAP was observed with the hybrid ACPs compared with u-ACP. The retarding effect of the Si or Zr species in the hybridized ACPs is probably due to these ions specifically blocking, by adsorption, potential sites for HAP nucleation and growth. The stability of ACP toward HAP conversion increased in the following order: u-ACP < Si-ACP < Zr-ACP. Hybrid ACP fillers, especially Zr-ACP, could be utilized in applications in which it is desired to enhance performance of composites, sealants, and/or adhesives in preventing demineralization or actively promoting remineralization.

Amorphous calcium phosphate.

ACP is a unique calcium phosphate in that it lacks long-range crystalline order. Yet the constancy in the composition of ACP over a wide range of solution conditions suggests a well-defined local structural unit. Although this order within disorder is the most distinguishing feature of ACP, the solution instability of ACP and its ready transformation into crystalline phases such as OCP and apatite may be of greater biological relevance. In particular, the initiating role ACP plays in matrix vesicle mineralization raises the importance of this phase from a mere laboratory curiosity to that of a key intermediate in skeletal calcification.

Physicochemical evaluation of bioactive polymeric composites based on hybrid amorphous calcium phosphates.

Amorphous calcium phosphate (ACP)-filled methacrylate composites were recently found to effectively remineralize in vitro caries-like enamel lesions. Their inferior mechanical properties compared to glass-filled composites, however, limit their use as a dental restorative material. In this study, the feasibility of introducing glass-forming elements (tetraethoxysilane or zirconyl chloride) during the low-temperature synthesis of ACP was investigated. Composites based on such hybrid fillers (mass fraction, 40%) were evaluated to establish whether hybridization strengthened the composites via improved interfacial interactions with the polymer phase without compromising the release of the mineral ions. Two types of visible-light cured resins were prepared: BTHZ resin from 2, 2-bis[p-(2'-hydroxy-3'-methacryloxypropoxy)phenyl]propane (BisGMA), triethylene glycol dimethacrylate (TEGDMA), 2-hydroxyethyl methacrylate (HEMA) and zirconyl methacrylate (ZrM), and TP resin from TEGDMA and pyromellitic glycerol dimethacrylate (PMGDM). Hybridized fillers and BTHZ- and TP-based composites were characterized by the IR spectroscopy, X-ray diffraction, dissolution/transformation kinetic studies, and biaxial flexure strength (BFS) testing before and after immersion in buffered saline solutions. The feasibility of improving the BFS via hybridization, while retaining, if not enhancing the remineralizing potential was demonstrated for BTHZ-based composites. Both BFS and remineralizing ability of the TP-composites, however, deteriorated upon their exposure to an aqueous environment. Therefore, hybridized ACP-filled BTHZ composites have a potential for utilization in more demanding restorative, sealant, and adhesive applications.

Anionic effects on the size and shape of apatite crystals grown from physiological solutions.

Comparatively little is known of the role tissue fluid electrolytes have in establishing the size and shape of apatite crystals deposited in skeletal tissues. In vitro accretion experiments using synthetic apatite seed crystals comparable in size to bone apatite were performed to assess the extent to which these crystalline features may be affected by direct electrolyte/mineral interactions. A constant composition method was used to maintain the accretion reactions under physiological-like solution conditions (1.33 mmol/liter Ca(2+), 1.0 mmol/liter total inorganic phosphate, (0 or 26) mmol/liter carbonate, 270 mmol/kg osmolality, pH 7.4, 37 degrees C). When the mass of the new accretions equaled the initial seed mass, the solids were harvested and the net change in crystal size resulting from the new accretions was assessed by X-ray diffraction line-broadening analysis. All the electrolytes examined in this study inhibited the accretion rate. The order of effectiveness was phosvitin > polyaspartate approximately polyglutamate > 1-hydroxyethylidene-1,1-bisphosphonate (HEBP) > bovine serum albumin (BSA) > citrate. Citrate and BSA also reduced the mean crystal size of the harvested solids compared with those harvested in the absence of added electrolyte, a finding that suggests that citrate and BSA suppressed growth of the seed crystals in favor of the proliferation of new smaller crystals. In contrast, a net increase in size following accretion compared with controls suggests that the other more strongly inhibiting electrolytes stimulated growth of the primary seed crystals and/or of the secondary crystals. These size changes, however, were anisotropic, with the anions effecting primarily increases in crystal width/thickness rather than in length. The effects were also more pronounced in the presence of carbonate. Our findings suggest that the strength of the interaction with the crystal surface may be relatively more important than molecular size or conformational complexity in establishing the effect that electrolytes have on apatite growth and proliferation. The results also suggest that adsorbed electrolytes may be a significant factor in controlling the size of apatite crystals in skeletal tissues by inducing proliferation of new crystals and/or affecting crystal shape by selectively modifying growth of the lateral dimensions.

The effect of fluoride on the size and morphology of apatite crystals grown from physiologic solutions.

In adult human bone, fluoride uptake is accompanied by an increase in apatite crystal size. This increase, however, is not isotropic but is restricted primarily to growth in width and/or thickness, with no measurable change in length. In the present study, seeded growth experiments were conducted in vitro to determine whether this anisotropic effect is physicochemical in origin, i.e., a direct result of F- selectively enhancing lateral crystal growth, or is an indirect consequence of F--induced alterations in cellular function and matrix development. The growth reactions were maintained at 37 degrees C under physiologic-like solution conditions (1.33 mmol/liter Ca2+, 1.0 mmol/liter total phosphate, 0 or 26 mmol/liter carbonate, 270 mmol/kg osmolality, pH 7.4) using constant-composition methods. When new accretions accumulated to three times the initial seed mass, the solids were collected and net crystal growth was assessed by X-ray diffraction line broadening analysis. The X-ray results revealed that the carbonate constituent in our physiologic-like solutions promoted the proliferation of new crystals at the expense of further growth of the seed apatite. Solution F- concentrations of approximately 2 micromol/liter partially offset the repressive effect that carbonate had on primary crystal growth. Moreover, F- stimulated seed crystal growth in the same anisotropic manner as had been observed for adult human bone apatite, a finding that suggests that the latter growth in vivo was the consequence, in part, of direct F--mineral interactions.

Mechanical properties of bioactive amorphous calcium phosphate/methacrylate composites.

OBJECTIVES: The aim of this study was to determine whether amorphous calcium phosphate (ACP)-containing composites, which have the ability to release mineralizing levels of Ca and PO4 ions, have appropriate mechanical properties for use as base and lining materials. METHODS: Composites of pyrophosphate-stabilized ACP particulates (mass fraction of 40%) and photo-activated methacrylate resins (mass fraction of 60%) were tested for biaxial flexure strength (BFS), diametral tensile strength (DTS), and compressive strength (CS). Hydroxyapatite (HAP; mass fraction of 40%), and micro-sized glass (mass fraction of 50%) composites as well as a commercial visible light curable base/liner were also tested. The significance between mean values was determined by Student-Newman-Keuls multiple comparisons (p < 0.05). RESULTS: BFS of dry and wet (24 h at 37 degrees C in water) ACP composites (60.3 and 62.0 MPa, respectively) were significantly lower than those of the comparison materials (79.2-109.3 MPa). CS values were likewise lower (62.9 MPa dry and 67.6 MPa wet vs 80.6-196.8 MPa) except for the wet base/liner (58.5 MPa). DTS of the dry ACP composite (21.8 MPa) was comparable with that of the HAP (22.8 MPa) and glass (25.5 MPa) composites, but lower than that of the base/liner (36.2 MPa). DTS decreased significantly when the ACP composite was wet (17.8 MPa). SIGNIFICANCE: These results suggest that the remineralizing ACP polymeric composites, although mechanically weaker in some respects than other polymeric composites, have properties suitable for use as base and lining materials.

Characterization of native and recombinant bone sialoprotein: delineation of the mineral-binding and cell adhesion domains and structural analysis of the RGD domain.

Bone sialoprotein is a small, sulfated, and phosphorylated integrin-binding glycoprotein apparently found only in tissues that eventually mineralize. Nondenatured bone sialoprotein (BSP) purified from rat osteosarcoma cell line (UMR 106-01 BSP) culture media is shown to have a hydroxyapatite Kd approximately 2.6 x 10(-9) M, perhaps the strongest affinity for this mineral of any of the matrix proteins. Both native BSP and a 47 kD fragment of UMR-BSP (Fragment 1 approximately 133A- approximately 265Y) are more potent inhibitors of seeded hydroxyapatite crystal growth than recombinant human BSP fragments lacking post-translational modifications. The recombinant proteins, however, do show reproducible inhibitory activity, suggesting that at least some of the strong mineral-binding properties are encoded directly within the protein sequence itself. BSP facilitates the adhesion of several cell types through its integrin binding (RGD) tripeptide sequence. Nuclear magnetic resonance (NMR) analysis of a 15N-enriched 59 amino acid recombinant domain containing the RGD tripeptide shows that the structure of this isolated domain is highly flexible with or without 5 mM calcium. Previous work has also shown that an endogenous fragment of UMR-BSP (Fragment 1) supports cell adhesion in the absence of the RGD sequence. In this report, non-RGD cell adhesion sites are localized within conserved amino- and carboxy-terminal tyrosine-rich domains of recombinant human BSP. Given the proximity of the latter non-RGD cell adhesion site to the RGD tripeptide, a model of BSP-receptor interactions is presented.

Quantitative assessment of the efficacy of amorphous calcium phosphate/methacrylate composites in remineralizing caries-like lesions artificially produced in bovine enamel.

Recent studies show that methacrylate-based composites with amorphous calcium phosphate (ACP) as a filler can release supersaturating levels of calcium and phosphate ions in proportions favorable for apatite formation. These findings suggest that such composites could be effectively used as coatings for remineralizing teeth damaged by tooth decay. To examine this hypothesis, we tested composites in vitro for their efficacy to remineralize artificially formed caries-like lesions in extracted bovine incisors. Single 120-microns-thick sagittal tooth sections were placed in holders that exposed only the carious enamel surface. The exposed surfaces were coated with a 1-mm- to 1.5-mm-thick layer of the composite containing, by mass, 40% apatite, silica, or P2O7(-4)-stabilized ACP and 60% photoactivated resin comprised of Bis-GMA, TEGDMA, HEMA, and ZrM. The photocured composite-coated sections were immersed either in a remineralizing solution for 4 weeks at 37 degrees C (static model) or cyclically immersed in demineralizing (0.5 h) and remineralizing solutions (11.5 h) for 2 weeks (dynamic model). Quantitative digital image analysis of matched 102 microns x 220 microns areas from contact microradiographs taken of the sections before and after immersion showed that lesions coated with ACP-filled composites fractionally recovered 71% +/- 33% of their lost mineral compared with 14% +/- 13% for apatite controls in the static model and 38% +/- 16% compared with -6% +/- 24% in the dynamic model. The results suggest that sealants based on ACP-filled methacrylate composites have the potential to remineralize carious enamel lesions.

Improved properties of amorphous calcium phosphate fillers in remineralizing resin composites.

OBJECTIVES: The rationale for this study was based on the hypothesis that the mechanical strength of methacrylate composites containing the bioactive filler, amorphous calcium phosphate, can be enhanced by synthesizing this filler in the presence of glass-forming agents. Specifically, this study was conducted to prepare composites with zirconia- and silica-modified amorphous calcium phosphate fillers, and to determine whether the remineralization potential from the release of calcium and phosphate ions and the mechanical properties of the corresponding methacrylate composites were enhanced. METHODS: The modified amorphous calcium phosphates were synthesized at pH 10.5 by mixing 800 mmol/L Ca(NO3)2 solutions and either 250 mmol/L zirconylchloride (ZrOCl2) or 4.4 mol/L tetraethoxysilane (TEOS) solutions with solutions containing 525 mmol/L Na2HPO4 and 11 mmol/L Na4P2O7. After washing and drying, the amorphous calcium phosphates were mixed with visible light-activated resins and photopolymerized to form composite disks that were then examined for their ability to release Ca2+ and total ionic phosphate (PO4(3-) + HPO4(2-) + H2PO4-, hereafter indicated as PO4) by immersion in HEPES-buffered (pH 7.4) saline at 37 degrees C. Solution ion concentrations were compared at regular intervals up to 265 h. Biaxial flexural strengths of the composites before and after immersion were compared, and significant differences were established by Student's test (p < 0.05). RESULTS: Both ZrOCl2- and TEOS-modified amorphous calcium phosphate composite disks released Ca2+ and PO4 ions at sustained levels requisite for remineralization to occur. The transformation of amorphous calcium phosphate into hydroxyapatite within the composites was also retarded, particularly in the case of amorphous calcium phosphate modified with ZrOCl2. Biaxial flexure strength values of composite disks showed that TEOS- and ZrOCl2-amorphous calcium phosphate-filled composites increased in strength by 33% and 21% before immersion and by 25% and 27% after immersion, respectively, compared to unmodified amorphous calcium phosphate composites (controls). All strength increases except TEOS after immersion were significant (p < 0.05). SIGNIFICANCE: Properly modified amorphous calcium phosphate fillers can be used to prepare bioactive composites with enhanced mechanical properties for more demanding dental applications without compromising their remineralizing potential.

Bladder calcifications after photodynamic therapy: analysis of a rare complication.

OBJECTIVES: We analyzed bladder calcifications occurring after photodynamic therapy administered for the treatment of superficial bladder cancer, a finding not previously reported after this treatment. METHODS: Bladder biopsies from 20 patients undergoing photodynamic therapy were evaluated. Bladder calcifications were identified in 2 patients and analyzed for composition. RESULTS: One patient had diffuse microcrystalline deposition in two biopsies composed of calcium oxalate monohydrate A. A second patient had a focal stone at a healing biopsy site composed of monoclinic calcium hydrogen phosphate dihydrate (brushite) (66%), calcium oxalate (25%), hydroxyapatite (6%), and protein (3%). CONCLUSIONS: Rare calcium oxalate and brushite calcifications were identified after photodynamic therapy and presumed to occur because of tissue injury associated with treatment.

In vitro inhibition of membrane-mediated calcification by novel phosphonates.

The effects of a series of novel phosphonates on the kinetics of mineral development in an ionophore-primed 7:2:1 phosphatidylcholine (PC): dicetylphosphate (DCP): cholesterol (Chol) liposomal model system are reported. When present at 2.5 micromol/liter or 25 micromol/liter concentrations in the solution surrounding the liposomes, the investigated phosphonates did not significantly delay the initial formation of hydroxyapatite-like calcium phosphate salts (HAP) within the liposomes or the penetration of HAP crystals through the enclosing membranes. However, the phosphonates variably retarded the subsequent growth and proliferation of the HAP crystals once they became directly exposed to the phosphonate-containing solution. The effectiveness of phosphonates in inhibiting extraliposomal precipitation strongly depended on their structure. The inhibitory action on active surface growth sites of released intraliposomal crystals was found to be the most effective if the phosphonate molecule contained two phosphonic groups linked to the same C atom. At a phosphonate concentration of 25 micromol/liter, the following general order of effectiveness was established: geminal bisphosphonate >/= geminal tetrakisphosphonate > bisacylphosphonates > monoacylphosphonate > bisalkylphosphonate. Within the bisacylphosphonate family, the highest inhibitory action was observed when four or five -CH2- groups separated the ketophosphonic groups.

Rapidly forming apatitic mineral in an osteoblastic cell line (UMR 106-01 BSP).

This study evaluated a rapid biomineralization phenomenon exhibited by an osteoblastic cell line, UMR 106-01 BSP, when treated with either organic phosphates [beta-glycerophosphate (beta-GP), Ser-P, or Thr-P], inorganic phosphate (P(i)), or calcium. In a dose-dependent manner, these agents (2-10 mM) stimulated confluent cultures to deposit mineral in the cell layer (ED50 of approximately 4.6 mM for beta-GP (30 +/- 2 nmol Ca2+/microgram DNA) and approximately 3.8 mM (29 +/- 2 nmol Ca2+/microgram DNA) for P(i)) with a plateau in mineral formation by 20 h (ET50 approximately 12-15 h). beta-GP or P(i) treatment yielded mineral crystals having an x-ray diffraction pattern similar to normal human bone. Alizarin red-S histology demonstrated calcium mineral deposition in the extracellular matrix and what appeared to be intracellular paranuclear staining. Electron microscopy revealed small, needle-like crystals associated with fibrillar, extracellular matrix deposits and intracellular spherical structures. Mineral formation was inhibited by levamisole (ED50 approximately 250 microM), pyrophosphate (ED50 approximately 1-10 microM), actinomycin C1 (500 ng/ml), cycloheximide (50 micrograms/ml), or brefeldin A (1 microgram/ml). These results indicate that UMR 106-01 BSP cells form a bio-apatitic mineralized matrix upon addition of supplemental phosphate. This process involves alkaline phosphatase activity, ongoing RNA and protein synthesis, as well as Golgi-mediated processing and secretion.

Effect of calcium ions on hydroxyapatite formation from the hydrolysis of anhydrous dicalcium phosphate.

Effects of solution calcium ions on hydroxyapatite (HAP) formation from the hydrolysis of dicalcium phosphate anhydrous (DCPA) were investigated under controlled solution conditions. The hydrolysis experiments (2.5 mmol DCPA/250 mL) were carried out at fixed pH in the absence or presence of CaCl2 (0-200 mmol/L) by standard pH stat techniques at 37 degrees C. The rate of hydrolysis was much faster, and the yield of HAP approximately doubled in the presence of CaCl2 when compared to no CaCl2, but no significant difference was observed over the range of Ca2+ concentrations examined (25-200 mmol/L). At equivalent conversion times, the crystallinity of the HAP formed was also greater in the presence of CaCl2. Calculations showed that much less DCPA dissolved to reach solution equilibrium with respect to HAP in the presence than in the absence of CaCl2. We conclude, therefore, that solution Ca2+ is an important factor in controlling HAP formation from the hydrolysis of acidic calcium phosphates such as DCPA.

Effect of 1-hydroxyethylidene-1,1-bisphosphonate on membrane-mediated calcium phosphate formation in model liposomal suspensions.

The bisphosphonate, 1-hydroxyethylidene-1,1-bisphosphonate (HEBP), was examined for its effect on calcium phosphate precipitation in pH 7.4, 22 degrees C suspensions of 7:2:1 PC:phosphatidylcholine (PC):dicetylphosphate (DCP):cholesterol (Chol) and 7:1:1 PC:phosphatidylserine (PS):Chol liposomes. HEBP (0.5-50 mumol/l) in the suspending medium had little, if any, effect on precipitation that formed inside phosphate-rich (50 mmol/l) aqueous interiors of liposomes as a result of ionophore (X-537A) driven 2.25 mmol/l Ca2+ influxes from the medium. On the other hand, HEBP had a significant negative impact on the subsequent spread of the precipitate into the surrounding medium when the latter was made metastable with 1.5 mmol/l total inorganic phosphate (PO4). The inhibitory effect of HEBP was more strongly felt in the 7PC:1PS:1Chol liposomal suspensions, with only 1 mumol/l HEBP needed to effectively block extraliposomal precipitation compared to 7.5 mumol/l for 7PC:2DCP:1Chol suspensions. Direct encapsulation of HEBP (1-1000 mumol/l) together with PO4 in the aqueous cores of 7PC:2DCP:1Chol liposomes reduced somewhat (approximately 30%) intraliposomal yields and delayed but did not block extraliposomal precipitate development. These results provide a possible physicochemical explanation for the suppression of matrix vesicle initiated mineralization in ectopically-induced osteoid tissue of HEBP treated mice [1]. In particular, the liposome results suggest that membrane phosphatidylserine interactions with mineral may enhance HEBP's effectiveness in vivo.

Effect of ultrafilterable fragments from chondroitinase and protease-treated aggrecan on calcium phosphate precipitation in liposomal suspensions.

A liposome-centered endogenous precipitation method was used to investigate the effect of ultrafilterable fragments from the enzymatic digestion of rat chondrosarcoma aggrecan on the formation of insoluble calcium phosphate salts in buffered solutions at pH 7.4 and 22 degrees C. Unlike the intact aggrecan and its major chondroitin sulfate and core protein components, disaccharide units from chondroitinase degradation of the aggrecan and small (< 3 kg/mol molecular weight) fragments from protease digestion of the core structure were found to be only weakly inhibitory toward mineral formation. Corresponding reductions in Ca(2+)-binding indicate that these fragments were unable to absorb to active sites on the apatite surface for long enough periods to significantly hinder crystal growth. The data suggest that controlled enzymatic breakdown of aggrecan may be one possible mechanism by which the calcification of growth plate cartilage is allowed to advance in vivo.

The effect of supersaturation on apatite crystal formation in aqueous solutions at physiologic pH and temperature.

The importance of supersaturation in the dynamics of apatite precipitation from aqueous solutions is well-established. To determine whether this parameter has a comparable impact on the concomitant development of the textural properties of this phase, such as crystal size and shape, we investigated mineral accretion in synthetic solutions seeded with 0.67 g/L apatite over a range of supersaturations at pH 7.4 and 37 degrees C. A dual specific-ion electrode-controlled titration method was used to maintain the seeded reactions under the following solution conditions: 1.0 to 1.8 mmol/L Ca2+, 0.67 to 1.2 mmol/L total phosphate (PO4), Ca/PO4 (initial) = 1.5, 143 mmol/L KNO3, and 10 mmol/L HEPES. Samples were collected for chemical and textural analyses when the seed apatite was reduced by new accretions to 1/2, 1/4, 1/8, 1/16, and 1/32 of the total solids in suspension. All new accretions were found to be apatitic. At the lowest supersaturation, accretion occurred primarily by growth of the seed crystals. However, at the highest supersaturation examined, the crystals at the end of the experiments were actually smaller, on average, than the original seeds, even though the total mass increased 32-fold. The results suggest that proliferation of new crystals supplanted growth of the seed crystals as supersaturation was increased. The results also suggest that differences in tissue fluid supersaturation may contribute to the large disparity in dimensions between dentin and enamel apatite crystals.