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Cell-loaded apatite microcarriers present as potential scaffolds for direct in-vivo delivery of cells post-expansion to promote bone regeneration. The objective of this study was to evaluate the osteogenic potency of human foetal mesenchymal stem cells (hfMSC)-loaded apatite microcarriers when implanted subcutaneously in a mouse model. This was done by examining for ectopic bone formation at 2 weeks, 1 month and 2 months, which were intended to coincide with the inflammation, healing and remodelling phases, respectively. Three histological examinations including haematoxylin and eosin staining to examine general tissue morphology, Masson's trichrome staining to identify tissue type, and Von Kossa staining to examine extent of tissue mineralisation were performed. In addition, immunohistochemistry assay of osteopontin was conducted to confirm active bone formation by the seeded hfMSCs. Results showed a high level of tissue organisation and new bone formation, with active bone remodelling being observed at the end of 2 months, and an increase in tissue density, organisation, and mineralisation could also be observed for hfMSC-loaded apatite microcarriers. Various cell morphology resembling that of osteoblasts and osteoclasts could be seen on the surfaces of the hfMSC-loaded apatite microcarriers, with presence of woven bone tissue formation being observed at the intergranular space. These observations were consistent with evidence of ectopic bone formation, which were absent in group containing apatite microcarriers only. Overall, results suggested that hfMSC-loaded apatite microcarriers retained their osteogenic potency after implantation, and provided an effective platform for bone tissue regeneration. Graphical Abstract: [ABSTRACT FROM AUTHOR]

The aim of the present study was to investigate the preferred orientation of biological apatite (BAp) as a new index of the quality of subchondral bone (SB) in knee joint osteoarthritis (OA). Ten OA and five normal knee joints were obtained. Thickness, quantity and bone mineral density (BMD) of SB were analyzed at the medial condyle of the femur in dry conditions by peripheral quantitative computed tomography. In addition, the preferred crystallographic orientation of the c-axis of BAp was evaluated as bone quality parameter using a microbeam X-ray diffractometer technique. BMD and thickness of SB were significantly increased in OA specimens compared to normal knee specimens (P < 0.01), and the preferred orientation of the c-axis of BAp along the normal direction of SB surface was significantly higher in OA specimens (P < 0.01), reflecting the change in stress of concentration in the pathological portion without cartilage. SB sclerosis in OA results in both proliferation of bone tissues and enhanced degree of preferential alignment of the c-axis of BAp. Our findings could have major implications for the diagnosis of clinical studies, including pathologic elucidation in OA. [ABSTRACT FROM AUTHOR]

This study examines the diet consumed by the population at the neighborhood center of Teopancazco, Teotihuacan, in the basin of Mexico. We used stable isotope analysis of δ13C and δ15N of the dentine collagen as well as the δ13C from enamel bioapatite from teeth (M2 and M3) of 39 of excavated burials. The results show that the diet consumed by the majority of the population during childhood and adolescence had high 13C enriched values in both dietary components: δ13C enamel_bioapatite = −2.0 ‰ ± 1.5 for the whole diet and δ13C dentine_collagen = − 9.4 ‰ ± 2.1 for the protein component. According to the bi-variable and multivariable models, some individuals consumed higher amounts of C4/CAM resources than those specified in cluster #2 (70 % C4 diet; ≥50 % C4/CAM protein). An inter-tissue adjustment of 2.3 ‰ was used to relate the enamel bioapatite data to the bone bioapatite of the models. Average δ15Ndentine_collagen was 10.2 ‰ ± 2.8 with high variability especially for four individuals, three of which belong to special burials, suggesting different access to meat or marine products due to their social or migration status and/or to pathological conditions. The data suggest that maize was the staple crop of the population not only at Teopancazco but for Teotihuacan as a whole, in the form of tortillas, tamales, atole, and protein from maize-fed animals with the inclusion of other CAM, C4, and C3 resources probably from the bountiful ecosystem of the basin of Mexico or from the wide trade system that Teotihuacan had with Mesoamerica. [ABSTRACT FROM AUTHOR]

Quantitative analysis of the orientational distribution of biological apatite (BAp) crystals is proposed as a new index of bone quality. This study aimed to analyze BAp c-axis orientation in ovariectomized (OVX) monkeys treated with amino-bisphosphonates minodronic acid and alendronate as reference. Sixty female monkeys aged 9-17 years were divided into five groups: one sham group and four OVX groups. The sham group and one OVX group were treated daily with vehicle for 17 months. The other three groups were treated daily with minodronic acid at doses of 0.015 and 0.15 mg/kg, and alendronate at 0.5 mg/kg orally, respectively. The seventh lumbar vertebrae were subjected to analysis of the preferential BAp c-axis orientation in the ventral cortical bone. The BAp c-axis orientation along the craniocaudal axis was significantly increased in the OVX monkeys. The high dose of minodronic acid suppressed the OVX-induced increase in the BAp c-axis orientation, whereas alendronate showed a non-significant tendency to suppress the increase in the orientation. In analysis with other parameters, the BAp c-axis orientation was positively correlated with bone formation indices in biochemical markers and bone histomorphometry and negatively correlated with the increase in lumbar bone mineral density. On the other hand, the BAp c-axis orientation was not correlated with bone resorption indices, except for the eroded surface. These results indicate that the increase in BAp c-axis orientation was ameliorated by minodronic acid treatment in OVX monkeys, mainly by suppression of bone formation increase. [ABSTRACT FROM AUTHOR]

The presence of infectious microorganisms in urinary stones is commonly inferred from stone composition, especially by the presence of struvite in a stone. The presence of highly carbonated apatite has also been proposed as a marker of the presence of bacteria within a stone. We retrospectively studied 368 patients who had undergone percutaneous nephrolithotomy (PCNL), and who also had culture results for both stone and urine. Urine culture showed no association with stone mineral content, but stone culture was more often positive in struvite-containing stones (73 % positive) and majority apatite stones (65 %) than in other stone types (54 %, lower than the others, P < 0.02). In 51 patients in whom the carbonate content of apatite could be measured, carbonate in the apatite was weakly predictive of positive stone culture with an optimal cutoff value of 13.5 % carbonate (sensitivity 0.61, specificity 0.80). In positive cultures of stones (all mineral types combined), organisms that characteristically produce urease were present in 71 % of the cases, with no difference in this proportion among different types of stone. In summary, the type of mineral in the stone was predictive of positive stone culture, but this correlation is imperfect, as over half of non-struvite, non-apatite stones were found to harbor culturable organisms. We conclude that mineral type is an inadequate predictor of whether a stone contains infectious organisms, and that stone culture is more likely to provide information useful to the management of patients undergoing PCNL. [ABSTRACT FROM AUTHOR]

Porous titanium and titanium alloys are promising scaffolds for bone tissue engineering, since they have the potential to provide new bone tissue ingrowth abilities and low elastic modulus to match that of natural bone. In the present study, porous Ti-7.5Mo alloy scaffolds with various porosities from 30 to 75 % were successfully prepared through a space-holder sintering method. The yield strength and elastic modulus of a Ti-7.5Mo scaffold with a porosity of 50 % are 127 MPa and 4.2 GPa, respectively, being relatively comparable to the reported mechanical properties of natural bone. In addition, the porous Ti-7.5Mo alloy exhibited improved apatite-forming abilities after pretreatment (with NaOH or NaOH + water) and subsequent immersion in simulated body fluid (SBF) at 37 °C. After soaking in an SBF solution for 21 days, a dense apatite layer covered the inner and outer surfaces of the pretreated porous Ti-7.5Mo substrates, thereby providing favorable bioactive conditions for bone bonding and growth. The preliminary cell culturing result revealed that the porous Ti-7.5Mo alloy supported cell attachment. [ABSTRACT FROM AUTHOR]

The kinetics of bone apatite crystallisation are examined using a novel approach to obtain quantitative, direction dependence features such as growth rate and activation energy. X-ray diffraction was employed for analysis of bovine, porcine and 'anorganic' bone specimens. Apatite coherence length was utilised as the independent variable within a Johnson-Mehl-Avrami (JMA) model. A direction averaged crystallisation activation energy of 183 ± 8 kJ mol was observed for the three bone groups. The Johnson-Mehl-Avrami ' n' exponent decreased with increasing temperature for all bone groups, indicating that apatite crystallisation changes to a diffusion limited process at higher temperatures. The results revealed little evidence to support any organic component 'protective' effect, and, on the contrary indicated that the organic matrix promotes apatite crystallisation. [ABSTRACT FROM AUTHOR]

The involvement of collagen in bone biomineralization is commonly admitted, yet its role remains unclear. Here we show that type I collagen in vitro can initiate and orientate the growth of carbonated apatite mineral in the absence of any other vertebrate extracellular matrix molecules of calcifying tissues. We also show that the collagen matrix influences the structural characteristics on the atomic scale, and controls the size and the three-dimensional distribution of apatite at larger length scales. These results call into question recent consensus in the literature on the need for Ca-rich non-collagenous proteins for collagen mineralization to occur in vivo. Our model is based on a collagen/apatite self-assembly process that combines the ability to mimic the in vivo extracellular fluid with three major features inherent to living bone tissue, that is, high fibrillar density, monodispersed fibrils and long-range hierarchical organization. [ABSTRACT FROM AUTHOR]

Vascular calcification, albeit heterogeneous in terms of biological and physicochemical properties, has been associated with ageing, lifestyle, diabetes, and chronic kidney disease (CKD). It is unknown whether or not moderately impaired renal function (CKD stages 2-4) affects the physiochemical composition and/or the formation of magnesium-containing tricalcium phosphate ([Ca,Mg][PO], whitlockite) in arterial microcalcification. Therefore, a high-resolution scanning X-ray diffraction analysis (European Synchrotron Radiation Facility, Grenoble, France) utilizing histological sections of paraffin-embedded arterial specimens derived from atherosclerotic patients with normal renal function ( n = 15) and CKD (stages 2-4, n = 13) was performed. This approach allowed us to spatially assess the contribution of calcium phosphate (apatite) and whitlockite to arterial microcalcification. Per group, the number of samples (13 vs. 12) with sufficient signal intensity and total lengths of regions (201 vs. 232 μm) giving rise to diffractograms ('informative regions') were comparable. Summarizing all informative regions per group into one composite sample revealed calcium phosphate/apatite as the leading mineral phase in CKD patients, whereas in patients with normal renal function the relative contribution of whitlockite and calcium phosphate/apatite was on the same order of magnitude (CKD, calcium phosphate/apatite 157 μm, whitlockite 38.7 μm; non-CKD, calcium phosphate/apatite 79.0 μm, whitlockite 94.1 μm; each p < 0.05). Our results, although based on a limited number of samples, indicate that chronic impairment of renal function affects local magnesium homeostasis and thus contributes to the physicochemical composition of microcalcification in atherosclerotic patients. [ABSTRACT FROM AUTHOR]

Water is well recognized as an important component in bone, typically regarded as a constituent of collagen, a pore-filling fluid in bone, and an adsorbed species on the surface of bone crystallites. The possible siting and role of water within the structure of the apatite crystallites have not been fully explored. In our experiments, carbonated hydroxyl- and fluorapatites were prepared in DO and characterized by elemental analysis, thermal gravimetric analysis, powder X-ray diffraction, and infrared and Raman spectroscopy. Two hydroxylapatites and two fluorapatites, with widely different amounts of carbonate were analyzed by solid state H NMR spectroscopy using the quadrupole echo pulse sequence, and each spectrum showed one single line as well as a low-intensity powder pattern. The relaxation time of 7.1 ms for 5.9 wt% carbonated hydroxylapatite indicates that the single line is likely due to rapid, high-symmetry jumps in translationally rigid DO molecules, indicative of structural incorporation within the lattice. Discrimination between structurally incorporated and adsorbed water is enhanced by the rapid exchange of surface DO with atmospheric HO. Moreover, a H resonance was observed for samples dried under a variety of conditions, including in vacuo heating to 150°C. In contrast, a sample heated to 500°C produced no deuterium resonance, indicating that structural water had been released by that temperature. We propose that water is located in the c-axis channels. Because structural water is observed even for apatites with very low carbonate content, some of the water molecules must lie between the monovalent ions. [ABSTRACT FROM AUTHOR]

New apatite (AP)/nanodiamond (ND) coating has been developed to improve physical and biological properties of stainless steel (SS) versus single AP coating. Homogeneously electrodeposited AP-ND layer demonstrates increased mechanical strength, interlayer cohesion and ductility. In the absence of serum, osteoblast-like MG63 cells attach well but poorly spread on both AP and AP-ND substrata. Pre-adsorption with serum or fibronectin (FN) improves the cellular interaction-an effect that is better pronounced on the AP-ND coating. In single protein adsorption study fluorescein isothiocyanate-labeled FN (FITC-FN) shows enhanced deposition on the AP-ND layer consistent with the significantly improved cell adhesion, spreading and focal adhesions formation (in comparison to SS and AP), particularly at low FN adsorption concentrations (1 μg/ml). Higher FN concentrations (20 μg/ml) abolish this difference suggesting that the promoted cellular interaction of serum (where FN is low) is caused by the greater affinity for FN. Moreover, it is found that MG63 cells tend to rearrange both adsorbed and secreted FN on the AP-ND layer suggesting facilitated FN matrix formation. [ABSTRACT FROM AUTHOR]

Bone is a hierarchically structured composite consisting of a protein phase (type I collagen) and a mineral phase (carbonated apatite). The objective of this study was to investigate the hierarchical structure of mineral in mature bone. A method to completely deproteinize bone without altering the original structure is developed, and the completion is confirmed by protein analysis techniques. Stereoscopy and field emission electron microscopy are used to examine the structural features from submillimeter- to micrometer- to nanometer-length scales of bovine femur cancellous bone. Stereoscopic images of fully deproteinized and demineralized bovine femur cancellous bone samples show that fine trabecular architecture is unaltered and the microstructural features are preserved, indicating the structural integrity of mineral and protein constituents. SEM revealed that bone minerals are fused together and form a sheet-like structure in a coherent manner with collagen fibrils. Well-organized pore systems are observed at varying hierarchical levels. Mineral sheets are peeled off and folded after compressive deformation, implying strong connection between individual crystallites. Results were compared with commercially available heat-deproteinized bone (Bio-Oss(®)), and evidence showed consistency in bone mineral structure. A two-phase interpenetrating composite model of mature bone is proposed and discussed. [ABSTRACT FROM AUTHOR]

In the present study, we fabricated magnesium doped apatite cement (md-AC) with rapid self-setting characteristic by adding the mixed powders of magnesium oxide and calcium dihydrogen phosphate (MO-CDP) into hydroxyapatite cement (HAC). The results revealed that the md-AC with 50 wt% MO-CDP could set within 6 min and the compression strength could reach 51 MPa after setting for 1 h, indicating that the md-AC had highly initial mechanical strength. The degradability of the md-AC in Tris-HCl solution increased with the increase of MO-CDP amount, and the weight loss ratio of md-AC with 50 wt% MO-CDP was 57.5 wt% after soaked for 12 weeks. Newly flake-like apatite could be deposited on the md-AC surfaces after soaked in simulated body fluid (SBF) for 7 days. Cell proliferation ratio of MG cells on md-AC was obviously higher than that of HAC on days 4 and 7. The cells with normal phenotype spread well on the md-AC surfaces and attached intimately with the substrate, and alkaline phosphatase (ALP) activity of the cells on md-AC significantly improved compared with HAC on day 7. The results demonstrate that the md-AC has a good ability to support cell proliferation and differentiation, and indicate a good cytocompatibility. [ABSTRACT FROM AUTHOR]

A simulated body fluid (SBF) with ion concentrations similar to body fluid, proposed by Kokubo et al., is widely used to evaluate bone-bonding potential through the formation of an apatite layer. To be confident of the evaluation of the potential for the apatite formation in SBF, standard substrates are required. Although Na2O–CaO–SiO2 glasses have been focused upon as candidate standard substrates, it has not been clarified whether the preparation conditions of the glasses affect their apatite formation potential in SBF. In this study, Na2O–CaO–SiO2 glasses were prepared by a conventional melting–quenching method with different melting periods and annealing processes to examine their properties, including apatite formation in SBF. The Na2O–CaO–SiO2 glasses show reproducible apatite-forming ability when prepared using moderate melting and annealing processes, and can be useful substrates to test the reproducibility of SBF. [ABSTRACT FROM AUTHOR]

Pathological calcification is common among for instance dialysis patients, and this causes an increase in mortality risk. An elevated serum phosphate concentration among those patients strongly correlates to this increase. In this work investigations of the conditions, composition, crystallinity and morphology of in vitro calcification are performed and related to results from in vivo studies. The study was performed under conditions mimicking physiological ones, i.e. a pH around 7.40, a temperature of 37°C, an ionic strength of 150 mM and ion concentrations close to those in human serum including the effects of elevated phosphate concentrations. The course of precipitation involves an initial precipitate that subsequently re-dissolves to give another precipitate, in accordance with the well-known Ostwald ripening theory. The final bulk precipitate consists of a macroscopically amorphous carbonated apatite. The amorphous apatite is formed from assemblies of spherical particles in the μm range, in turn composed of nano-crystalline needles of about 10 × 100 nm. Even the initially formed precipitate, as well as a small amount of precipitate that occurs on the liquid surface, consist of a carbonated calcium phosphate. The in vitro observed carbonated apatite bears strong resemblance to in vivo cardiovascular calcification known from literature. [ABSTRACT FROM AUTHOR]

The article provides information on the chemical composition and structure of bone minerals. It mentions that the solid phase in bones is identified as a crystalline calcium phosphate which is similar to geological apatite and reveals that there are important chemical and structural differences found in bone and skeletal tissues. It relates that other research have pointed that fractions of phosphate ions are found on the surfaces of bone minerals.

Apatite and brushite kidney stones share calcium and phosphate as their main inorganic components. We tested the hypothesis that these stone types differ in the amount of proteins present in the stones. Intact stones were intensively analyzed by microcomputed tomography (micro CT) for both morphology (including the volume of voids, i.e., space devoid of X-ray dense material) and mineral type. To extract all proteins present in kidney stones in soluble form we developed a three-step extraction procedure using the ground stone powder. Apatite stones had significantly higher levels of total protein content and void volume compared to brushite stones. The void volume was highly correlated with the total protein contents in all stones ( r 2 = 0.61, P < 0.0001), and brushite stones contained significantly fewer void regions and proteins than did apatite stones (3.2 ± 4.5% voids for brushite vs. 10.8 ± 11.2% for apatite, P < 0.005; 4.1 ± 1.6% protein for brushite vs. 6.0 ± 2.4% for apatite, P < 0.03). Morphological observations other than void volume did not correlate with protein content of stones, and neither did the presence or absence of minor mineral components. Our results show that protein content of brushite and apatite stones is higher than that was previously thought, and also suggest that micro CT-visible void regions are related to the presence of protein. [ABSTRACT FROM AUTHOR]

The influence of thin carbonate-containing apatite (CA) coating on trabecular bone response to cylindrical titanium fiber mesh (porosity of 85%, pore size of 200–300 μm, 2.8 mm diameter × 6 mm length) implants was investigated. Thin CA coatings were deposited by the so-called molecular precursor method. Molecular precursor solution was obtained by adding dibutylammonium diphosphate salt to Ca–EDTA/amine ethanol solution by adjusting Ca/P = 1.67. Sintered cylindrical titanium fiber mesh was immersed into molecular precursor solution and then tempered at 600 °C for 2 h. The immersion and tempering process was repeated three times. An adherent thin CA film could be deposited on the inside of titanium fiber mesh. After the immersion of a CA-coated titanium fiber mesh in simulated body fluid, apatite crystals precipitated on the titanium fiber mesh. Uncoated and CA-coated titanium fiber mesh was inserted into the trabecular bone of the left and right femoral condyles of rabbits. Histological and histomorphometrical evaluation revealed a significantly greater amount of bone formation inside the porous area of the CA-coated titanium fiber mesh after 12 weeks of implantation. The present results suggested that a thin CA-coated titanium mesh has better osteoconductivity and will be useful for a three-dimensional scaffold. [ABSTRACT FROM AUTHOR]

A versatile method for preparing spherical, micro- and macroporous (micro: 2–10 and macro: 150–550 μm pores), carbonated apatitic calcium phosphate (Ap-CaP) granules (2–4 mm in size) was developed by using NaCl crystals as the porogen. The entire granule production was performed between 21 and 37 °C. A CaP cement powder, comprising α-Ca3(PO4)2 (61 wt.%), CaHPO4 (26%), CaCO3 (10%) and precipitated hydroxyapatite, Ca10(PO4)6(OH)2 (3%), was dry mixed with NaCl crystals varying in size from 420 μm to 1 mm. Cement powder (35 wt.%) and NaCl (65 wt.%) mixture was kneaded with an ethanol–Na2HPO4 initiator solution, and the formed dough was immediately agitated on an automatic sieve shaker for a few minutes to produce the spherical granules. Embedded NaCl crystals were then leached out of the granules by soaking them in deionized water. CaP granules were micro- and macroporous with a total porosity of 50% or more. Granules were composed of carbonated, poorly crystallized, apatitic CaP phase. These were the first spherical and porous CaP granules ever produced from a self-setting calcium phosphate cement. The granules reached their final handling strength at the ambient temperature through the cement setting reaction, without having a need for sintering. [ABSTRACT FROM AUTHOR]

The surface transformation reactions of bioactive ceramics were studied in vitro in standard K9-SBF solution and in human blood serum (HBS)—containing simulated body fluid (SBF). The calcium phosphate ceramics used for this study were stoichiometric hydroxyapatite (HA), β-tricalcium phosphate (β-TCP) and brushite. Immersion of each calcium phosphate tested in this study, in simulated body fluid, led to immediate surface precipitation of apatite. The use of HBS resulted in a delay in the onset of precipitation and a significant inhibition of the dissolution reaction normally observed for brushite in solution. However, apatite formation still occurred. The use of HBS and SBF in this investigation, which has shown the ability to induce similar crystal growth as that observed in vivo, suggests that there is scope for the use of serum proteins in simulated body fluid in order to create a protein-rich surface coating on biomedical substrates. [ABSTRACT FROM AUTHOR]

The effects of heat treatment time and temperature on the delivery behaviour of Ibuprofen from hydroxyapatite particles were investigated in this study. The drug release was seen to follow Fickian diffusion for the initial period of release for all heat treatment conditions. The gradient of Fickian release increased with (1) increasing crystallite size, attributed to the decreasing amount of boundary area, and (2) with decreasing surface area, due to the reduction in porosity and hence tortuosity within the apatite particles. This study has shown that altering the heat treatment conditions used to calcine hydroxyapatite may alter its drug delivery abilities, whereby calcination temperature was noted to influence the drug release behaviour to a greater extent than calcination time. [ABSTRACT FROM AUTHOR]

We have reported that low-crystalline porous calcite block, which is useful as a bone substitute or a source material to prepare apatite-type bone fillers could be fabricated by exposing calcium hydroxide compact to carbon dioxide gas saturated with water vapor. In the present study, we investigated the effect of molding pressure on the transformation of calcium hydroxide into calcite and the mechanical strength of the carbonated compact. Transformation into calcite was almost completed within 72 h, however, a small amount of Ca(OH)2 still remained unreacted at higher molding pressure because of incomplete penetration of CO2 gas into the interparticle space due to dense packing of Ca(OH)2 particles. On the other hand, high molding pressure resulted in an increase in diametral tensile strength (DTS) of the calcite compact formed. Critical porosity of the calcite block was calculated as approximately 68%. [ABSTRACT FROM AUTHOR]

Computer modelling techniques were employed to investigate the effect of surface silanol groups on the strength of adhesion of apatite thin films to silica surfaces. To this end, we have studied a series of silica surfaces with different silanol densities and calculated their interaction with apatite thin films. Our findings indicate that apatite does not attach strongly to surface hydroxy groups, but that apatite should deposit at dehydrated silica surfaces, especially when the surface silicon and oxygen species rearrange to form O–Si–O links. Any dangling silicon and oxygen bonds at the silica surfaces are saturated by coordination to oxygen and calcium atoms in the apatite layer, but the extra reactivity afforded by these under-coordinated surface species does not necessarily lead to more favourable substrate/film interactions. The lowest energy silica/apatite interfaces are those where an undistorted apatite layer can be deposited on a regular, stable substrate surface. Our simulations support the suggestion, that in vivo surface hydroxy groups are first condensed to form O–Si–O bridges before deposition and growth of apatite. [ABSTRACT FROM AUTHOR]

Comparison of natural marine (nodular and granular) carbonate-apatites and pathogenic mineral structures formed in human cardiac valves by methods of scanning electron and high-resolution transmission microscopy revealed common morphological features corresponding to the bacterial origin of calcium phosphates. These features may be considered typomorphic ones regardless of their formation environments (in vivo or in nature). [ABSTRACT FROM AUTHOR]

High-resolution solid-state 19F NMR method has been used to examine the chemical structural forms of fluorine introduced into the bone tissue of experimental animals (Vistar rats) exposed to 30 day fluoride inhalation. It is shown that in the bone tissue three structural forms of fluorides are deposited: a solid phase of F-apatite and mobile nanoparticles of CaF2 and MgF2 (or KMgF3) with the ratio of fluorine concentration in these three forms ∼2:2:1. During the following 30 day rehabilitation this ratio remains constant, the total fluorine content in the bone tissue decreases ∼3 times and the F-apatite phase transforms into disordered (F, OH)-apatite. A protective effect of the zeolitic enterosorbent (klinoptilolite) on fluorine binding in the intoxication process was found, as well as the promotional effect of this enterosorbent on fluorine excretion during the postfluoride rehabilitation. [ABSTRACT FROM AUTHOR]

Human bone is a three-dimensional composite structure consisting of inorganic apatite crystals and organic collagen fibers. An attractive strategy for fabricating mimics of these types of composite biomaterials is to selectively grow apatite on polymers with control of structure, mechanical properties, and function. In the present study, silk/apatite composites were prepared by growing apatite on functionalized nanodiameter silk fibroin fibers prepared by electrospinning. The functionalized fibers were spun from an aqueous solution of silk/polyethylene oxide (PEO) (78/22 wt/wt) containing poly(L-aspartate) (poly-Asp), which was introduced as an analogue of noncollageous proteins normally found in bone. Silk fibroin associated with the acidic poly-Asp and acted as template for mineralization. Apatite mineral growth occurred preferentially along the longitudinal direction of the fibers, a feature that was not present in the absence of the combination of components at appropriate concentrations. Energy dispersive spectroscopy and x-ray diffraction confirmed that the mineral deposits were apatite. The results suggest that this approach can be used to form structures with potential utility for bone-related biomaterials based on the ability to control the interface wherein nucleation and crystal growth occur on the silk fibroin. With this level of inorganic-organic control, coupled with the unique mechanical properties, slow rates of biodegradation, and polymorphic features of this type of proteins, new opportunities emerge for utility of biomaterials. [ABSTRACT FROM AUTHOR]

Understanding the interaction of bioactive coatings with aqueous media is essential for development of systems possessing rapid osteointegration and durability. An in vitro study of a commercial, plasma sprayed hydroxyapatite coating has been undertaken. The coating behaviour when test coupons were immersed in water, simulated body fluid and foetal calf serum has been examined. The principal aim was to characterise, in detail, any structural changes to the coatings and, in particular, examine features of any new layers formed. The amorphous phase of the coating showed preferential dissolution in all media. The rate of dissolution was greatest in water and the process was initially retarded in the foetal calf serum. A nanocrystallite apatite layer was shown to precipitate on the coatings in all media although this was significantly enhanced in simulated body fluid. The features of this layer (e.g., lattice parameters, crystallite size etc.) were quantified by adopting a novel approach to the X-ray diffraction data analysis. The results are discussed in the context of similar studies and implications for in vivo behaviour. [ABSTRACT FROM AUTHOR]

In calcium renal stones, calcium oxalate and calcium phosphate in various crystal forms and states of hydration can be identified. Calcium oxalate monohydrate (COM) or whewellite and calcium oxalate dihydrate (COD) or weddellite are the commonest constituents of calcium stones. Calcium oxalate stones may be pure or mixed, usually with calcium phosphate or sometimes with uric acid or ammonium urate. The aim of this study was to compare the clinical and urinary patterns of patients forming calcium stones of different composition according to infrared spectroscopic analysis in order to obtain an insight into their etiology. The stones of 84 consecutive calcium renal stone formers were examined by infrared spectroscopy. In each patient, a blood sample was drawn and analysed for serum biochemistry and a 24-h urine sample was collected and analysed for calcium, phosphate, oxalate, citrate and other electrolytes. We classified 49 patients as calcium oxalate monohydrate (COM) stone formers, 32 as calcium oxalate dihydrate (COD) stone formers and three as apatite stone formers according to the main component of their stones. Patients with COM stones were significantly older than patients with COD stones ( P<0.002). Mean daily urinary calcium and urinary saturation with respect to calcium oxalate were significantly lower in patients with COM than in those with COD stones ( P<0.000). Patients with calcium oxalate stones containing a urate component (≤10%) presented with higher saturation ( P<0.012) with respect to uric acid in their urine (and lower with respect to calcium oxalate and calcium phosphate, respectively P<0.024 and P<0.003) in comparison with patients without a urate component in the stone. Patients with calcium oxalate stones with a calcium phosphate component (≥15%) showed higher ( P<0.0016) urinary saturation levels with respect to calcium phosphate (and lower with respect to uric acid ( P<0.009), compared with patients forming stones without calcium phosphate or with a low calcium phosphate component. Patients with calcium stones mixed with urate had a significantly lower urinary pH ( P<0.002) and urinary calcium ( P<0.000), and patients with calcium phosphate >15%, higher urinary pH ( P<0.004) and urinary calcium ( P<0.000). In conclusion, in the evaluation of the individual stone patient, an accurate analysis of the stone showing its exact composition and the eventual presence of minor components of the stone is mandatory in order to plan the correct prophylactic treatment. Patients with “calcium stones” could require various approaches dependent on the form and hydration of the calcium crystals in their stones, and on the presence of “minor” crystalline components that could have acted as epitaxial factors. [ABSTRACT FROM AUTHOR]

The chemical and phase composition of pathogenic minerals formed on human heart valves (cardiolytes) has been studied. It is shown that cardiolytes are organomineral aggregates, whose mineral component is represented by nonstoichiometric carbonate-containing hydroxyl apatites with a variable $\{\backslash rm\; Ca\}\; /\; \{\backslash rm\; P\}$ atomic ratio. [ABSTRACT FROM AUTHOR]

Previous 31P cross-polarization and differential cross-polarization magic angle spinning (CP/MAS and DCP/MAS) solid-state NMR spectroscopy studies of native bone and of the isolated crystals of the calcified matrix synthesized by osteoblasts in cell culture identified and characterized the major PO4-3 phosphate components of the mineral phase. The isotropic and anisotropic chemical shift parameters of the minor HPO4-2 component in bone mineral and in mineral deposited in osteoblast cell cultures were found to differ significantly from those of brushite, octacalcium phosphate, and other synthetic calcium phosphates. However, because of in vivo and in vitro evidence that phosphoproteins may play a significant role in the nucleation of the solid mineral phase of calcium phosphate in bone and other vertebrate calcified tissues, the focus of the current solid-state 31P NMR experiments was to detect the possible presence of and characterize the phosphoryl groups of phosphoproteins in bone at the very earliest stages of bone mineralization, as well as the possible presence of calcium-phosphoprotein complexes. The present study demonstrates that by far the major phosphate components identified by solid-state 31P NMR in the very earliest stages of mineralization are protein phosphoryl groups which are not complexed with calcium. However, very small amounts of calcium-complexed protein phosphoryl groups as well as even smaller, trace amounts of apatite crystals were also present at the earliest phases of mineralization. These data support the hypothesis that phosphoproteins complexed with calcium play a significant role in the initiation of bone calcification. [ABSTRACT FROM AUTHOR]

Atomic force microscopy (AFM) was used to obtain three-dimensional images of isolated mineralites extracted from young postnatal bovine bone. The mean mineralite size is 9 nm × 6 nm × 2.0 nm, significantly shorter and thicker than the mineralites of mature bovine bone measured by the same technique. Mineralites of the young postnatal bone can be accommodated within the hole zone regions of a quasi-hexagonally packed collagen fibril in the fashion described by Hodge [9] in which laterally adjacent hole zone regions form continuous “channels” across the diameter of a fibril for a distance of at least 10 nm. Deposition of mineralites of the size noted above in this void volume of the fibrils would result in little or no distortion of the collagen molecules or supramolecular structure of the collagen fibril. The new AFM data supporting this claim is consistent with findings obtained by electron microscopy and low-angle x-ray and neutron diffraction that mineralites formed within collagen fibrils during initial stages of calcification occur within the hole zone region. However, the deposition of additional mineralites in the intermolecular spaces between collagen molecules in the overlap region of the fibrils would significantly distort the fibrils since the space available between adjacent molecules is considerably less than even the smallest dimension of the mineralites. [ABSTRACT FROM AUTHOR]

The assembly of amelogenin protein into nanospheres is postulated to be a key factor in the stability of enamel extracellular matrix framework, which provides the scaffolding for the initial enamel apatite crystals to nucleate and grow. Adsorption isotherms were evaluated in order to investigate the nature of interactions of amelogenin nanospheres with hydroxyapaite crystals in solution, where their assembly status and particle size distribution are defined. We report that the adsorption isotherm of a recombinant mouse amelogenin (rM179) on synthetic hydroxyapatite crystals can be described using a Langmuir model indicating that amelogenin nanospheres adsorb onto the surface of apatite crystals as binding units with defined adsorption sites. The adsorption affinity and the maximum adsorption sites were 19.7 × 105 L/mol and 6.09 × 10-7 mol/m2, respectively, with an r2 value of 0.99. Knowing the composition and particle size distribution of amelogenin nanospheres under the condition of adsorption experiments, we have calculated the number of nanospheres and the crystal surface area covered by each population of nanospheres at their maximum adsorption. It was found that total maximum binding covers 64% of the area unit. This observation supports the speculation that amelogenin binding onto apatite surface is selective and occurs only at certain sites. [ABSTRACT FROM AUTHOR]

Fractures of the orbital floor often need to be reconstructed with rigid materials that provide a stable orbital floor and reestablish the correct orbital volume. In addition to autologous bone grafts, many alloplastic materials are available such as hydroxyapatite and, recently, expanded polytetrafluoroethylene reinforced with fluorinated ethylene propylene also known as reinforced SAM Gore-Tex. This study compared the efficacy and biocompatibility of reinforced and nonreinforced SAM Gore-Tex to those of hydroxyapatite and autogenous bone graft for reconstructing the orbital floor. A defect measuring 1.5×1.0 cm was created in the left orbital floor in 24 short-hair cats while the right side served as the control. The cats were divided into four groups, the defect being reconstructed with an autogenous iliac crest corticocancellous bone graft (group A), hydroxyapatite (group B), nonreinforced Gore-Tex (group C), or reinforced Gore-Tex (group D). Computed tomography was performed postoperatively at 1 and 8 weeks, and at the conclusion of the study specimens from the left orbit were harvested to evaluate the histological biocompatibility of each material. Reinforced SAM Gore-Tex possessed excellent physical properties. It was malleable yet firm enough and therefore easily contoured during repair of the defect while strong enough to support the orbital contents. It could even be sutured to the orbital rim or a fixating plate to ensure further stabilization, if necessary. Additionally, the material demonstrated exemplary biocompatibility as it initiated a minimal inflammatory response. As healing progressed, a growth of host tissue into the material was noted in all histological specimens, thereby augmenting the stability of the implant and reducing the incidence of extrusion. [ABSTRACT FROM AUTHOR]

Peritubular dentin (PTD) is a relatively dense mineralized tissue surrounding tooth dentin tubules, whose composition and mode of formation are still unclear. Fourier transform infrared microspectroscopic studies of the organic and mineral components of the highly developed horse PTD indicate that the peritubular matrix is less abundant than the intertubular matrix but is also mainly composed of collagen, which is more hydrated. These data suggest that most of the crystals are located outside the collagen fibrils and probably not associated with protein components. The crystals in PTD have nearly the same crystallinity as those in intertubular spaces, showing comparable amounts of carbonate ions, although some PO4 groups have different nonapatitic environments. Horse PTD composition is very similar to that of ITD, with collagen as the main protein component and carbonated apatite as the mineral fraction but the different proportions of these constituents and the greater water content in PTD suggest a different organization. [ABSTRACT FROM AUTHOR]

The mechanical properties of bone are studied mostly for reasons related to skeletal pathology. However, bone is also very interesting from a material science perspective because it is a natural hierarchical composite material. The mechanical properties of bone depend on both the structural arrangement and the properties of the constituting materials, namely the organic polymer collagen and the inorganic salt apatite. While the mechanical properties of bone samples at the macroscopic scale are measured routinely, mechanical tests on micrometer-sized specimens are still at development stage. In this paper, protocols for measuring the elasticity of cancellous bone trabeculae are reviewed. The published values for the elastic modulus of trabeculae vary between 1 GPa and 15 GPa. Reasons for this broad range of values may be located in the intrinsic difficulties of preparing, handling, and testing inhomogeneous, anisotropic and asymmetric micro-samples. We discuss the major error sources in existing testing procedures and suggest potential strategies to enhance their performance. [ABSTRACT FROM AUTHOR]

Raman spectroscopy has been used to demonstrate, for the first time, that calcium mineralization in the core of the major lateral teeth of the chiton Acanthopleura echinata takes place as an ordered process, with crystalline carbonated apatite being the first mineral deposited. Deposition begins at the top of the tooth core, under the so-called tab region, progresses down the interior surface of the tab and lepidocrocite layer, and then extends outwards to the anterior surface. Mineralization is not initiated until the lepidocrocite layer has isolated the core of the tooth from the magnetite cap. The last region to be infiltrated is the anterior basal region of the tooth cusp, immediately above the junction zone. The junction zone is also a region of high ion density, as determined by energy dispersive spectroscopy (EDS) analysis, but we show here for the first time that it is free of mineral deposits, acting instead as a transfer and storage region. [ABSTRACT FROM AUTHOR]

Bisphosphonates have been widely used in the treatment of human bone pathologies including osteoporosis. In this case, bisphosphonates have been shown to reduce bone resorption, thereby increasing the mass and mechanical resistance of bone. Determining the effects of these molecules on the properties of the bone apatite crystals could provide a better insight into the mechanism of bisphosphonate/bone interaction. The aim of this study was to determine the ultrastructural effects of a third generation bisphosphonate (tiludronate) on the morphology, size, distribution, chemical composition, and structure of apatite crystals in bone (trabecular) in a rat osteoporotic model. Four groups of rats were studied: (1) sham operated, (2) untreated ovariectomized (OVX), (3) OVX rats which received 35 mg/kg of tiludronate, (4) OVX rats which received 160 mg/kg of tiludronate. The rats of groups 3 and 4 received tiludronate orally in 2 consecutive days every week for 1 year. Scanning electron microscopy (SEM), high and low resolution transmission electron microscopy (TEM), and electron microprobe analysis (EDX) were used for the ultrastructural characterization of the bone mineral. This study demonstrated that tiludronate slightly increased the width of bone apatite crystals without changing any other crystal characteristics. [ABSTRACT FROM AUTHOR]