Calcium Phosphate Crystallites Research Articles

Solid-State Nuclear Magnetic Resonance Identifies Abnormal Calcium Phosphate Formation in Diseased Bones.

The crystallites of calcium phosphate (CaP) in bones consist of hydroxyl apatite (HA) and amorphous calcium phosphate (ACP). These nanoscale structures of CaP are sculptured by biological bone formation and resorption processes and are one of the crucial factors that determine the overall strength of the constructs. We used one- and two-dimensional 1H-31P solid-state nuclear magnetic resonance (SSNMR) to investigate the nanoscopic structural changes of CaP. Two quantitative measurables are deduced based on the heterogeneous linewidth of 31P signal and the ratio of ACP to HA, which characterize the mineral crystallinity and the relative proportion of ACP, respectively. We analyzed bones from different murine models of osteopetrosis and osteoporosis and from human samples with osteoporosis and osteoarthritis. It shows that the ACP content increases notably in osteopetrotic bones that are characterized by defective osteoclastic resorption, whereas the overall crystallinity increases in osteoporotic bones that are marked by overactive osteoclastic resorption. Similar pathological characteristics are observed for the sclerotic bones of late-stage osteoarthritis, as compared to those of the osteopetrotic bones. These findings suggest that osteoclast-related bone diseases not only alter the bone density macroscopically but also lead to abnormal formation of CaP crystallites. The quantitative measurement by SSNMR provides a unique perspective on the pathology of bone diseases at the nanoscopic level.

Pathological calcification in osteoarthritis: an outcome or a disease initiator?

In the progression of osteoarthritis, pathological calcification in the affected joint is an important feature. The role of these crystallites in the pathogenesis and progression of osteoarthritis is controversial; it remains unclear whether they act as a disease initiator or are present as a result of joint damage. Recent studies reported that the molecular mechanisms regulating physiological calcification of skeletal tissues are similar to those regulating pathological or ectopic calcification of soft tissues. Pathological calcification takes place when the equilibrium is disrupted. Calcium phosphate crystallites are identified in most affected joints and the presence of these crystallites is closely correlated with the extent of joint destruction. These observations suggest that pathological calcification is most likely to be a disease initiator instead of an outcome of osteoarthritis progression. Inhibiting pathological crystallite deposition within joint tissues therefore represents a potential therapeutic target in the management of osteoarthritis.

Effectiveness of Calcium Phosphate Desensitising Agents in Dental Hypersensitivity Over 24 Weeks of Clinical Evaluation.

Background: Calcium phosphate-based compounds are used to treat dental hypersensitivity (DH). Their long-term clinical behaviour needs further research. This study compared the 24-week effectiveness of Teethmate Desensitizer (TD), a pure tetracalcium phosphate (TTCP) and dicalcium phosphate dihydrate (DCPD) powder/water, to that of Dentin Desensitizer (DD), and Bite & White ExSense (BWE), both of calcium phosphate crystallites. Methods: A total of 105 subjects were selected. A random table was utilised to form three groups of 35 subjects. DH was evaluated using the evaporative sensitivity, tactile sensitivity tests, and the visual analogue scale (VAS) of pain. Response was recorded before the application of the materials (Pre-1), immediately after (Post-0), at 1 week (Post-1), 4 weeks (Post-2), 12 weeks (Post-3) and 24 weeks (Post-4). The non-parametric distribution was assessed with the Shapiro–Wilk statistical test. Intra-group differences for the six time points were evaluated with the Friedman statistical test and the Kruskal–Wallis test. Results: All the materials decreased DH after 24 weeks in comparison to Pre-1. However, the TTCP/DCPD cement showed the greatest statistical efficiency. Conclusions: The significant decrease of VAS scores produced by TD in the long term suggest the material as the most reliable in the clinical relief of DH.

Optimized biological tools: ultrastructure of rodent and bat teeth compared to human teeth

Enamel, the outer layer of the teeth of vertebrates, is the hardest tissue in their body. In general, enamel is highly mineralized and has a special ultrastructure that directs the mechanical properties of teeth and thereby their biological functions. However, there are differences between the enamel ultrastructures of different species. The authors compare the teeth of the Eurasian beaver Castor fiber, the African mole rat Fukomys kafuensis and the common pipistrelle bat Pipistrellus pipistrellus by high-resolution analytical methods, including scanning electron microscopy, quantitative wavelength-dispersive X-ray spectroscopy and X-ray powder diffraction. The enamel of all animals consists of long, thin and differently oriented calcium phosphate crystallites (length > 1 µm; thickness about 50 nm). On the outer surface of the beaver tooth, a thin layer of brown iron oxide was found. The diameter of the dentin tubuli was between 1 and 3 µm for all species – that is, comparable to that for human teeth.

Influence of calcium and aluminum on crystallization of vivianite, [formula omitted

The iron(II) phosphate vivianite, Fe3(PO4)2·8H2O, has been precipitated at 37 °C from solution of ammonium iron(II) sulfate (Mohr’s salt) and ammonium phosphate, to which a small amount of calcium nitrate or aluminum potassium sulfate (alum) was added. The relatively high temperature was chosen to minimize the formation of crystal aggregates, which would otherwise complicate study of morphology. Crystallization kinetics was determined by pH recording, and the resulting crystals were studied by polarization microscopy. The main effect of Al observed was formation of small, irregular crystals. In contrast, Ca had little or no effect on morphology, but in three cases at intermediate additive concentrations kinetics changed from the usual combination of spiral growth and surface nucleation to pure spiral growth. It is proposed that the screw dislocations leading to growth spirals originate either from calcium phosphate crystallites acting as heteronuclei or from nuclei containing both Fe and Ca and with structure deviating from that of pure vivianite.

Calcium phosphate compound formed on electrospun chitosan nanofibers using modified simulated body fluid

In the present work, chitosan (CS) nanofibers were mineralized in order to enhance their osteoconductivity and the calcium phosphate (CaP) compounds formed on the surface were characterized. CS nanofibers blended with CS and polyvinyl alcohol were fabricated using an electrospinning technique. These fibers were soaked in a modified simulated body fluid (m-SBF) to encourage the formation of CaP for 1–7 days. Sphere-like CaP crystallites with average diameter of 350 nm were evenly formed on the surface of CS nanofibers. Their density increased with increasing soaking time in m-SBF. These crystallites were aggregated with the nano-sized crystallites which had plate-like shape and low crystallinity. High-resolution transmittance electron microscope image showed that these crystallites of CaP had β-tricalcium phosphate crystalline structure which had higher solubility compared with octacalcium phosphate or hydroxyapatite. The mineralized CaP/CS nanocomposite fibers in m-SBF could be promising biodegradable scaffolds for bone tissue regeneration.

Preparation and fabrication studies of three dimensionally ordered nano-, micro- and meso-scale calcium phosphate crystallites scaffold for artificial bone materials (3-DOMm)

Preparation and fabrication studies of three dimensionally ordered nano-, micro- and meso-scale calcium phosphate crystallites scaffold for artificial bone materials (3-DOMm)

Intracellular pathway and subsequent transformation of hydroxyapatite nanoparticles in the SAOS-2 osteoblast cell line.

Internalization of hydroxyapatite nanoparticles in SAOS-2 osteoblasts for 2 and 24 h was investigated in vitro using 5 and 50 µg/mL nanoparticles in culture medium. No cytotoxic effects were observed in a PrestoBlue viability assay. Focused ion beam-scanning electron microscopy and transmission electron microscopy were used to study nanoparticle trafficking inside cells and to characterize the physicochemical properties of the remodeled nanoparticles. Nanoparticles were actively internalized by cells and maintained in intracellular membrane-bound compartments. Dissolution of hydroxyapatite nanoparticles was observed inside phagolysosome in all samples. After 24 h of internalization in cell culture assays, reprecipitation of calcium phosphate minerals was observed in membrane-bound compartments in 5 and 50 µg/mL samples. Compared to the original nanoparticles, the reprecipitated calcium phosphate phase presented a different morphology, structure, and chemical composition. Two sample preparation methods were used and confirmed that reprecipitation of the calcium phosphate crystallites occurred in the intracellular environment and not during electron microscopy sample preparation. Reprecipitation of calcium phosphate prevented the release of large amounts of calcium and phosphate ions inside the cells. This phenomenon may be linked to physiological processes in the cell that control the concentration and trafficking of intracellular calcium ions, which are highly controlled by cells. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 106A: 428-439, 2018.

Use of Wet Chemical Method to Prepare β Tri-Calcium Phosphates having Macro- and Nano-crystallites for Artificial Bone

Calcium phosphate crystallites were prepared by wet chemical method for use in artificial bone. In order to obtain β-tricalcium phosphate (TCP), nano-crystalline calcium phosphate (CaP) was precipitated at 37℃ and at pH5.0±0.1 under stirring using highly active Ca(OH)₂ in DI water and an aqueous solution of H3PO4. The precipitated nano-crystalline CaP solution was kept at 90℃ for the growth of CaP crystallites. Through the growing process of CaP crystallites, we were able to obtain various sizes of rectangular CaP crystallites according to the crystal growing times. Dry nano-crystalline CaP powders at 37℃ were mixed with dry macro-crystalline CaP crystallites and the shaped mixture sample was fired at 1150℃ to make a β-TCP block. Several tens of nm powders were uniformly coated on the surface, which was comprised of powders of several tens of μm, using a vibrator. The mixing ratio between the nanometer powders and the micrometer powders greatly affected the mechanical strength of the mixture block; the most appropriate ratio of these two materials was 50 wt% to 50 wt%. The sintered block showed improved mechanical strength, which was caused by the solid state interaction between the nano-crystalline β-TCP and the macro-crystalline β-TCP.

Zinc-Containing Restorations Create Amorphous Biogenic Apatite at the Carious Dentin Interface: A X-Ray Diffraction (XRD) Crystal Lattice Analysis

The aim of this research was to assess the ability of amalgam restorations to induce amorphous mineral precipitation at the caries-affected dentin substrate. Sound and caries-affected dentin surfaces were subjected to both Zn-free and Zn-containing dental amalgam restorations. Specimens were submitted to thermocycling (100,000 cycles/5°C-55°C, 3 months). Dentin surfaces were studied by atomic force microscopy (nanoroughness), X-ray diffraction, field emission scanning electron microscopy, and energy-dispersive analysis, for physical and morphological surface characterization. Zn-containing amalgam placement reduced crystallinity, crystallite size, and grain size of calcium phosphate crystallites at the dentin surface. Both microstrain and nanoroughness were augmented in caries-affected dentin restored with Zn-containing amalgams. Caries-affected dentin showed the shortest mineral crystallites (11.04 nm), when Zn-containing amalgams were used for restorations, probably leading to a decrease of mechanical properties which might favor crack propagation and deformation. Sound dentin restored with Zn-free amalgams exhibited a substantial increase in length of grain particles (12.44 nm) embedded into dentin crystallites. Zn-containing amalgam placement creates dentin mineralization and the resultant mineral was amorphous in nature. Amorphous calcium phosphate provides a local ion-rich environment, which is considered favorable for in situ generation of prenucleation clusters, promotong further dentin remineralization.

Randomized, placebo-controlled study of the efficacy of a calcium phosphate containing paste on dentin hypersensitivity

ObjectiveHypersensitivity of non-carious cervical lesions (DH) is a frequently encountered disease. This randomized, controlled, single-blind crossover study evaluated the effectiveness of a calcium phosphate containing desensitizer paste (TAP) on DH in comparison to water as placebo (PLA). MethodsIn this clinical trial 35 patients were randomly assigned to the test and the negative control group. Using a 10cm long VAS (visual analog scale) patients should respond with DH score >6 on one tooth in each of two quadrants for allocation. Pain stimuli were a 2-seconds air blast (AB) and probe scratching (PS) of the exposed dentin. VAS scores were determined pre-operatively (PRE), immediately after treatment (POST), at 1 week, 1, 3 and finally after 6 months. ResultsBoth TAP and PLA applications decreased DH significantly at POST and throughout the 6-months recalls (p<0.001). Pain reductions upon AB stimulation of TAP treated teeth, assessed at POST and 6 months were 35 and 55%, upon PS stimuli 21 and 54%, respectively. PLA treated lesions responded to AB at POST and after 6 months with 20 and 36% pain reduction, to PS with 11 and 30% pain reduction, respectively. Differences between TAP and PLA pain scores were statistically significant at all recalls (p<0.05). SignificanceTAP paste reduced DH successfully during this 6-months trial. The calcium phosphate crystallites included in the paste and the presumed hydroxyapatite precipitates upon exposure to saliva were hypothetically able to occlude open dentinal tubules, at least to some extent. TAP is considered a biocompatible desensitizer paste.

Component, Mobility and Specific Surface Area of Urinary Crystallites with Different Sizes in Urine of Healthy Human and Lithogenic Patients

The property difference of urine crystallites with different sizes in the urines of calcium oxalate (CaOx) calculi patients and healthy controls was comparatively investigated. Nanoparticle size analyzer was used to study the mobility of urine crystallites. Fast Fourier transformation (FFT) and and energy dispersive X-ray spectroscopy (EDS) was performed to detect the components of urinary crystallites. The specific surface area (Sg) of nanoscale urinary crystallites is greater than that of micron-grade urinary crystals. The mobility of the controls was more negative than that of the calculi patients. The effect of particle size on mobility was greater than that of the charges on surface of the particles. The amount of small-sized urine crystallites in the healthy subjects was significantly higher than that of the patients. However, the number of large micron-scale crystals in the controls was reduced remarkably. These results showed that the formation of CaOx calculi was closely related to the presence of UA and calcium phosphate crystallites in urine. The urinary system of calculi patients is unstable and very susceptible to urine crystallite aggregation.

Understanding Nano-Anatomy of Healthy and Carious Human Teeth: a Prerequisite for Nanodentistry

The anatomy of human teeth reflects its usage. Spatially resolved X-ray scattering permits quantitative studies of the characteristic arrangement of the anisotropic calcium phosphate crystallites and the collagen fibers within the hard tissues of the crown. The present study summarizes the distinctive nanometer-sized anatomical features of the tooth hard tissues including their interface taking advantage of spatially resolved synchrotron radiation-based small-angle X-ray scattering. The comparison of slices from eight teeth indicates a long-range organization of tooth nanostructures.

Experimental Investigation on Mechanism of Hydrophilic Acrylic Intraocular Lens Calcification

To construct a model simulating intraocular lens (IOL) opacification attributable to the formation of calcium phosphate deposits and to investigate the kinetics of deposit formation. Prospective laboratory investigation. Department of Ophthalmology, Medical School and Department of Chemical Engineering, Laboratory of Inorganic and Analytical Chemistry, University of Patras, Greece. Three hydrophilic acrylic IOLs (26% water content) were placed inside a 10-mL double-walled thermostated reactor simulating the anterior chamber. Simulated aqueous humor was injected continuously into the reactor using a pump with variable speed. The observation of IOLs was carried out in situ daily by optical microscopy. Scanning electron microscopy and energy-dispersive radiographic spectroscopy were used for the identification of the morphologic features and the composition of the deposits. The lenses were removed and inspected 5, 9, and 12 months after the initiation of the experiment. Investigation showed deposits of calcium phosphate crystallites in the interior of opacified IOLs. However, these deposits were not observed on the surface of the IOLs. In agreement with earlier reports by our group and in the literature, IOL opacification is the result of calcification. It is suggested that the surface hydroxyl groups of the polyacrylic polymeric components of the IOLs are capable of inducing surface nucleation and crystal growth of calcium phosphates. However, most important is the finding that the calcification of IOLs is initiated from their interior through the development of sufficiently high local supersaturation, realized through the diffusion of calcium and phosphate ions.

Double-Hydrophilic Block Copolymers as Precipitation Inhibitors for Calcium Phosphate and Iron(III)

Novel double-hydrophilic block copolymers, maleic anhydride (MA)−allylpolyethoxy carboxylate (APEC) and maleic anhydride (MA)−ammonium allylpolyethoxy sulfate (APES), were specially designed and synthesized from allyloxy polyethoxy ether (APEO) to inhibit the precipitation of calcium phosphate and iron (III). Structures of APEO, APEC, MA−APEC, APES, and MA−APES were characterized by FT-IR. The study shows that both MA−APEC and MA−APES have significant ability to inhibit the precipitation of calcium phosphate at the dosage of 6 mg/L, approximately showing 99 and 90% inhibition, respectively. The data of the light transmittance of ferrous solutions show that, compared to MA−APES, MA−APEC has superior ability to stabilize iron (III) in solutions. The light transmittance of ferrous solutions is about 23% in the presence of MA−APEC when the dosage is 6 mg/L, whereas in the presence of MA−APES, it is about 31% at a dosage of 12 mg/L. Scanning electron microscopy (SEM) shows that the copolymers influence the morphology of calcium phosphate crystallites. Transmission electron microscopy (TEM) indicates the excellent inhibition results from the formation of core−shell structure. Double-hydrophilic block copolymers of MA−APEC and MA−APES have also been proven to be effective inhibitors of calcium phosphate and iron (III) even at elevated solution temperature, pH, and Ca2+ and Fe2+ concentration.

&lt;i&gt;In Situ&lt;/i&gt; Growth of Carbon Nanotubes in Hydroxyapatite Matrix by Chemical Vapor Deposition

Carbon nanotubes (CNTs)/hydroxyapatite (HA) nanocomposites have been successfully fabricated by a novel method for the biomedical applications, which is in situ growing CNTs in HA matrix in a chemical vapor deposition (CVD) system. The results show that it is feasible to in situ grow CNTs in HA matrix by CVD for the fabrication of CNTs/HA nanocomposites. Multi-walled CNTs with 50-80 nm in diameter have been grown in situ from HA matrix with the pretreatment of sintering at 1473K in air. The nanocomposites are composed with carbon crystals in CNTs form, HA crystallites and calcium phosphate crystallites, one of most important CaP bioceramics. And the CNTs content is about 1% proportion by weight among the composites in our experiments, which can enhance the HA mechanical properties and the CNTs content does not affect the HA performances. These CNTs/HA nanocomposites have the potential application in the biomedical fields.

A new evaporation-based method for the preparation of biomimetic calcium phosphate coatings on metals

This study reports a new method to prepare biomimetic calcium phosphate coatings on titanium, stainless steel, CoCrMo, and tantalum. The method does not require surface etching, high supersaturation, or tight control of solution conditions. Metallic samples were dipped into a supersaturated calcium phosphate solution, withdrawn, and left to dry at room temperature. Calcium phosphate crystallites formed on and completely covered the surfaces by repeating the dip-and-dry treatment. The crystallite-covered surfaces readily grew to calcium phosphate coatings when immersed in the supersaturated solution. The mechanism of the treatment was suggested to be an evaporation-induced surface crystallization process.

Accelerating calcium phosphate growth on NaOH-treated poly-(lactic- co-glycolic acid) by evaporation-induced surface crystallization

Poly(lactic- co-glycolic acid) (PLGA) is a promising material for the regeneration of bone tissue, but its surface properties are not optimal for the application. Coating the surface of PLGA with a continuous layer of calcium phosphate is an effective approach to address the limitation. Current coating techniques for PLGA require immersion in supersaturated calcium phosphate solutions for days to weeks. In this study, we report a simple technique to accelerate the coating process to only 2 h immersion in supersaturated solutions. PLGA pellets were first treated with NaOH to increase their hydrophilicity. The NaOH-treated PLGA pellets were repeatedly dipped in a supersaturated calcium phosphate solution and dried in air. After 10 times of the dip-and-dry treatment, a layer of calcium phosphate crystallites uniformly covered the surfaces of the pellets. After the crystallite-covered pellets were immersed in the supersaturated solution for 2 h, about 5-μm thick continuous calcium phosphate coatings formed on the surfaces. The dip-and-dry technique was also applied on a variety of metals and porous structures. An evaporation-induced surface crystallization process was suggested as the mechanism for the dip-and-dry treatment.

Opacification of Hydrophilic Acrylic Intraocular Lens Attributable to Calcification: Investigation on Mechanism

To identify the nature and to investigate the biochemical mechanisms leading to late opacification of implanted hydrophilic acrylic intraocular lenses (IOLs). Retrospective laboratory investigation. setting: Department of Ophthalmology, Medical School, Department of Chemical Engineering, Laboratory of Inorganic and Analytical Chemistry, University of Patras and FORTH-ICEHT, Greece. study population: Thirty IOLs were explanted one to 12 years postimplantation attributable to gradual opacification of the lens material. observation procedures: Materials analysis was done using scanning electron microscopy (SEM) equipped with a microanalysis probe (EDS), confocal microscopy, x-ray diffraction (XRD), and Fourier transform infrared (FTIR) for the identification of the substances involved in the opacified lenses. SEM investigation showed plate-like as well as prismatic nanoparticle deposits of calcium phosphate crystallites on the surface and in the interior of opacified IOLs. The plate-like deposits exhibited morphology and particle size typical for octacalcium phosphate (OCP), while the respective characteristics of the prismatic nanocrystals were typical of hydroxyapatite (HAP). EDS analysis confirmed the chemical composition of the deposits. Aqueous humor analysis showed that the humor is supersaturated with respect to both OCP and HAP, favoring the formation of the thermodynamically more stable HAP, while the formation and kinetic stabilization of other transient phases is also very likely. In vitro experiments using polyacrylic materials confirmed the clinical findings. Hydrophilic acrylic IOLs' opacification may be attributed to the deposition of calcium phosphate crystallites. HAP is the predominant crystalline phase of these crystallites. Surface hydroxyl groups of the polyacrylic materials facilitate surface nucleation and growth.

Thermal behaviour of composites aluminosilicate-calcium phosphates

A new type of aluminosilicate matrix calcium phosphate crystallites composites (ACPC) was synthesized and studied for osseous bone applications. The room temperature synthesis of the aluminosilicate matrix and composites was described. Thermal treatments of compounds allowed the adaptability of some parameters (pH, porosity and mechanical properties). Structure of heat treated composites were characterized by XRD and FTIR. The influence of thermal treatment on the mechanical properties, the porosity and the pH was studied for two temperatures (250 and 500°C). Results evidenced the ability to control the pH, the high level of porosity (»70%) and the good mechanical properties, allowing to consider that ACPC are potential biomaterials for osseous bone application.