The Use of Scanning Probe Microscopy to Investigate Crystal-Fluid Interfaces

Journal of Crystal Growth

Habelitz

2011

a b s t r a c tHuman dental enamel forms over a period of 2-4 years by substituting the enamel matrix, a protein gel mostly composed of a single protein, amelogenin with fibrous apatite nanocrystals. Self-assembly of amelogenin and the products of its selective proteolytic digestion are presumed to direct the growth of apatite fibers and their organization into bundles that eventually comprise the mature enamel, the hardest tissue in the mammalian body. This work aimed to establish the physicochemical and biochemical conditions for the growth of apatite crystals under the control of a recombinant amelogenin matrix (rH174) in combination with a programmable titration system. The growth of apatite substrates was initiated in the presence of self-assembling amelogenin particles. A series of constant titration rate experiments was performed that allowed for a gradual increase of the calcium and/or phosphate concentrations in the protein suspensions. We observed a significant amount of apatite crystals formed on the substrates following the titration of rH174 sols that comprised the initial supersaturation ratio equal to zero. The protein layers adsorbed onto the substrate apatite crystals were shown to act as promoters of nucleation and growth of calcium phosphates subsequently formed on the substrate surface. Nucleation lag time experiments have showed that rH174 tends to accelerate precipitation from metastable calcium phosphate solutions in proportion to its concentration. Despite their mainly hydrophobic nature, amelogenin nanospheres, the size and surface charge properties of which were analyzed using dynamic light scattering, acted as a nucleating agent for the crystallization of apatite. The biomimetic experimental setting applied in this study proves as convenient for gaining insight into the fundamental nature of the process of amelogenesis.

Section: Resultssupporting

confidence: 90%

Amelogenin as a promoter of nucleation and crystal growth of apatite

Journal of Crystal Growth

Habelitz

2011

“…First of all, HAp is typified by incongruent dissolution, releasing more Ca 2+ than PO 4 3− in the acidic milieu and more PO 4 3− than Ca 2+ under alkaline conditions 62 . Conversely, formation of phosphate network is the first step in nucleation of HAp, the reason for which high phosphate and low calcium concentrations favor HAp formation more than the other way around 63 and are routinely used in biomimetic mineralization experiments 64 . As a result, the activation barriers for Ca 2+ and H x PO 4 x-3 ions differ and the crystal growth rate can be twice higher at high [H x PO 4 x-3 ]/[Ca 2+ ] ratios, even when the supersaturation ratio is the same 65 .…”

Section: Resultsmentioning

confidence: 99%

One ion to rule them all: the combined antibacterial, osteoinductive and anticancer properties of selenite-incorporated hydroxyapatite

Iyer

2017

J. Mater. Chem. B

Although hydroxyapatite (HAp) has been doped with dozens of different ions, the quest for an ion imparting a combination of properties conducive to bone healing is still ongoing. Because of its protean potency and the similarity in size and shape to the phosphate tetrahedron, selenite ion presents a natural ionic substitute in HAp. The incorporation of selenite into synthetic HAp using two different methods – co-precipitation and ion-exchange sorption - was studied for its effect on crystal properties and on a triad of biological responses: antibacterial, anticancer and osteoinductive. Co-precipitation yielded HAp with higher selenite contents than sorption and the stoichiometry of HAp richest in selenite was represented as Ca9.75(PO4)5.75(SeO3)0.25(OH)1.75. Crystallinity of HAp decreased in direct proportion with the amount of selenite incorporated. Because of their lower selenite content, HAp powders prepared by ion-exchange exhibited a consistently higher crystallinity compared to the co-precipitated ones. Annealing partially recovered the crystallinity, yet the difference in crystallinity between powders prepared by co-precipitation and by ion-exchange remained, suggesting that the amorphization is mainly due to structural incorporation of selenite, not its effect on the crystal growth kinetics. The addition of selenite changed the morphology of HAp nanoparticles from acicular to rounded and affected the crystal lattice parameters in different ways depending on whether the powders were annealed or not. As for the annealed powders, the incorporation of selenite contracted the lattice in both a and c crystallographic directions. In the agar diffusion assay, the effectiveness of HAp was more dependent on the presence or absence of selenite in it than on its concentration and was highest against E. coli and S. aureus, moderately high against S. enteritidis and ineffective against P. aeruginosa. In liquid inoculation tests, on the other hand, the antibacterial activity of HAp was directly proportional to the amount of selenite contained in it. The viability of K7M2 osteosarcoma cells decreased in direct proportion with the amount of selenite in HAp and was significantly different from the untreated control and from pure HAp at contents equal to or higher than 1.9 wt.%. In contrast, no reduction was observed in the viability of primary fibroblasts treated with HAp incorporating different amounts of selenite ions, suggesting their potentially selective anticancer activity: lethal for the cancer cells and harmless for the healthy cells. Finally, mRNA expression of bone gamma-carboxyglutamate protein (BGLAP3) was higher in differentiated MC3T3-E1 osteoblastic cells treated with selenite-incorporated HAp particles than in cells treated with pure HAp. The osteoinductive effect was due to an overall higher metabolic activity of cells treated with the particles and not due to increased proliferation. In such a way, a triad of antibacterial, osteoinductive and anticancer activities was attributed to selenite-incorporated ...

“…Namely, Ca 2+ ions are more mobile than H x PO 4 x-3 ions, which tend to reside on surfaces longer and present the starting points for nucleation of CPs 59,60 . Per the Ostwald-Lussac rule, the growth of a most stable phase proceeds through successive growth and dissolution of less stable ones in the direction spanning from the least to the most stable.…”

Section: Discussionmentioning

confidence: 99%

Nonlinear oscillatory dynamics of the hardening of calcium phosphate bone cements

Rau

2017

RSC Adv.

Here we report on the nonlinear, oscillatory dynamics detected in the evolution of phase composition during the setting of different calcium phosphate cements, two of which evolved toward brushite and one toward hydroxyapatite as the final product. Whereas both brushite-forming cements contained ion-doped β-tricalcium phosphate as the initial phase, the zinc-containing one yielded scholzite as an additional phase during setting and the oscillations between these two products were pronounced throughout the entire 80 h setting period, long after the hardening processes was over from the mechanical standpoint. Oscillations in the copper-containing system involved the amount of brushite as the main product of the hardening reaction and they progressed faster toward an equilibrium point than in the zinc-containing system. Initially detected with the use of in situ energy-dispersive X-ray diffractometry, the oscillations were confirmed with a sufficient level of temporal matching in an in situ Fourier transform infrared spectroscopic analysis. The kinetic reaction analysis based on the Johnson-Mehl-Avrami-Kolmogorov model indicated an edge-controlled nucleation mechanism for brushite. The hydroxyapatite-forming cement comprised gelatin as an additional phase with a role of slowing down diffusion and allowing the detection of otherwise rapid oscillations in crystallinity and in the amount of the apatitic phase on the timescale of minutes. A number of possible causes for these dynamic instabilities were discussed. The classical chemical oscillatory model should not apply to these systems unless in combination with less exotic mechanisms of physicochemical nature. One possibility is that the variations in viscosity, directly affecting diffusion and nucleation rates and accompanying growth and transformation from the lower to the higher interfacial energy per the Ostwald-Lussac rule, are responsible for the oscillatory dynamics. The conception of bone replacement materials and tissue engineering constructs capable of engaging in the dynamics of integration with the natural tissues in compliance with this oscillatory nature may open a new avenue for the future of this type of medical devices. To succeed in this goal, the mechanism of these and similar instabilities must be better understood.