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Although the measurement of tooth quality is necessary for precise prediction of caries formation, typical measurement methods include tooth-hardness measurements and absorption spectroscopy, which generally use infrared light irradiation. These methods are destructive or invasive, and obtaining two-dimensional information in the oral cavity is difficult. Mid-infrared emissions from the surface of an object reflect intrinsic vibrations of molecules in the object. In this study, a mid-infrared passive spectroscopic imaging system was developed using an inexpensive uncooled microbolometer array sensor with an optimized multi-slit, which eliminated the cancellation of interference intensities between two adjacent emission points, to obtain two-dimensional information from an object without external infrared light irradiation. First, the feasibility of obtaining two-dimensional information on tooth quality using the proposed system was examined, and emission spectra attributed to phosphate ions in hydroxyapatite (HAp), the main component of enamel, were successfully obtained from bovine teeth. Further, the hardness of bovine teeth was measured, and a correlation ( R 2 = 0.8067) between the Vickers hardness and peak area ratio of phosphate ions assigned to the crystalline and amorphous phases of a tooth was established. Additionally, tooth-hardness visualization in a non-contact manner was demonstrated as two-dimensional information using the obtained regression equation.

Spectral Photon Counting Computed Tomography (SPCCT), a ground-breaking development in CT technology, has immense potential to address the persistent problem of metal artefacts in CT images. This study aims to evaluate the potential of Mars photon-counting CT technology in reducing metal artefacts. It focuses on identifying and quantifying clinically significant materials in the presence of metal objects. A multi-material phantom was used, containing inserts of varying concentrations of hydroxyapatite (a mineral present in teeth, bones, and calcified plaque), iodine (used as a contrast agent), CT water (to mimic soft tissue), and adipose (as a fat substitute). Three sets of scans were acquired: with aluminium, with stainless steel, and without a metal insert as a reference dataset. Data acquisition was performed using a Mars SPCCT scanner (Microlab 5×120); operated at 118 kVp and 80 μA. The images were subsequently reconstructed into five energy bins: 7-40, 40-50, 50-60, 60-79, and 79-118 keV. Evaluation metrics including signal-to-noise ratio (SNR), linearity of attenuation profiles, root mean square error (RMSE), and area under the curve (AUC) were employed to assess the energy and material-density images with and without metal inserts. Results show decreased metal artefacts and a better signal-to-noise ratio (up to 25%) with increased energy bins as compared to reference data. The attenuation profile also demonstrated high linearity (R2 >0.95) and lower RMSE across all material concentrations, even in the presence of aluminium and steel. Material identification accuracy for iodine and hydroxyapatite (with and without metal inserts) remained consistent, minimally impacting AUC values. For demonstration purposes, the biological sample was also scanned with the stainless steel volar implant and cortical bone screw, and the images were objectively assessed to indicate the potential effectiveness of SPCCT in replicating real-world clinical scenarios., Competing Interests: NO authors have competing interests., (Copyright: © 2024 Tariq et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.)

Electron spin resonance coupled with uranium-series dating (ESR/U-series) of carbonate hydroxyapatite in tooth enamel is the main technique used to obtain age determinations from Pleistocene fossils beyond the range of radiocarbon dating. This chronological information allows to better understand diachronic change in the palaeontological record, especially with regard to the evolution of the genus Homo. Given the relative paucity of human teeth at palaeontological and archaeological localities, ESR/U-series is widely applied to the teeth of ungulate species. However, the accuracy of ESR/U-series ages is greatly affected by the incorporation of uranium in the enamel during burial in sediments. It has been shown that uranium content is positively correlated with an increased degree of atomic order in carbonate hydroxyapatite crystals, the latter determined using infrared spectroscopy. Here we present a reference infrared spectral library of tooth enamel from African ungulates, based on the grinding curve method, which serves as baseline to track the diagenetic history of carbonate hydroxyapatite in different species and thus select the best-preserved specimens for dating., (© 2024. The Author(s).)

Teeth are considered unique as fingerprints for identification purposes. Their structure and resilience mean they can remain for thousands and millions of years withstanding extreme conditions, including burning. During burning, bones undergo carbonization at approximately 400°C and calcination at approximately 700°C. This study aimed to investigate the effects of carbonization and calcination on dental tissue. It involved nondestructive analyses of 58 extracted human teeth before and after burning, using x-ray diffraction, micro-CT, and high-resolution confocal microscopy. The results revealed that during carbonization, dentin volume decreased in two thirds of the sample, accompanied by crack formation and significant reduction in hydroxyapatite crystal size (p<0.001). During calcination, dentin volume decreased in all teeth, along with a significant deepening of the cracks (p<0.001), while enamel crystal size increased slightly. Initial changes in teeth occurred at lower temperatures than had once been assumed, as indicated by the cracks during carbonization, and there was up to a 36% decrease in dentin volume during calcination, which should be considered when measuring burnt teeth. The results of this research provide new insight into understanding dental tissue response to burning. Thus, dental remains may contribute to the knowledge needed to reconstruct anthropological and forensic scenarios involving burning., Competing Interests: Declaration of Competing Interest None, (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: In recent years, natural bone extracellular matrix (ECM)-inspired materials have found widespread application as scaffolds for bone tissue engineering. However, the challenge of creating scaffolds that mimic natural bone ECM's mechanical strength and hierarchical nano-micro-macro structures remains. The purposes of this study were to introduce an innovative bone ECM-inspired scaffold that integrates a 3D-printed framework with hydroxyapatite (HAp) mineralized graphene oxide-collagen (GO-Col) microscaffolds and find its application in the repair of mandibular bone defects., Methods: Initially, a 3D-printed polycaprolactone (PCL) scaffold was designed with cubic disks and square pores to mimic the macrostructure of bone ECM. Subsequently, we developed multi-layer mineralized GO-Col-HAp microscaffolds (MLM GCH) to simulate natural bone ECM's nano- and microstructural features. Systematic in vitro and in vivo experiments were introduced to evaluate the ECM-inspired structure of the scaffold and to explore its effect on cell proliferation and its ability to repair rat bone defects., Results: The resultant MLM GCH/PCL composite scaffolds exhibited robust mechanical strength and ample assembly space. Moreover, the ECM-inspired MLM GCH microscaffolds displayed favorable attributes such as water absorption and retention and demonstrated promising cell adsorption, proliferation, and osteogenic differentiation in vitro. The MLM GCH/PCL composite scaffolds exhibited successful bone regeneration within mandibular bone defects in vivo., Conclusions: This study presents a well-conceived strategy for fabricating ECM-inspired scaffolds by integrating 3D-printed PCL frameworks with multilayer mineralized porous microscaffolds, enhancing cell proliferation, osteogenic differentiation, and bone regeneration. This construction approach holds the potential for extension to various other biomaterial types., (© 2024. The Author(s).)

Ameloblasts are specialized cells derived from the dental epithelium that produce enamel, a hierarchically structured tissue comprised of highly elongated hydroxylapatite (OHAp) crystallites. The unique function of the epithelial cells synthesizing crystallites and assembling them in a mechanically robust structure is not fully elucidated yet, partly due to limitations with in vitro experimental models. Herein, we demonstrate the ability to generate mineralizing dental epithelial organoids (DEOs) from adult dental epithelial stem cells (aDESCs) isolated from mouse incisor tissues. DEOs expressed ameloblast markers, could be maintained for more than five months (11 passages) in vitro in media containing modulators of Wnt, Egf, Bmp, Fgf and Notch signaling pathways, and were amenable to cryostorage. When transplanted underneath murine kidney capsules, organoids produced OHAp crystallites similar in composition, size, and shape to mineralized dental tissues, including some enamel-like elongated crystals. DEOs are thus a powerful in vitro model to study mineralization process by dental epithelium, which can pave the way to understanding amelogenesis and developing regenerative therapy of enamel., (© 2023. The Author(s).)

This study aimed to analyze the crystalline structure of dental enamel in pediatric patients with chronic kidney disease (CKD) by X-ray diffraction (XRD). The six tested samples had a mineral composition similar to hydroxyapatite, according to sheet JCPDS(Joint Committee on Powder Diffraction Standards) card #09-0432, which is normally found in dentine, and presented a lower amount of whitlockites (Ca, Mg) 3 (PO 4 ) 2 . Pattern phases showed an increase in organic matter and a decrease in inorganic matter. At an interval of approximately 2θ = 15.7° to 27.2°, amorphous organic matter corresponding to hydrated glucose was found. The hydroxyapatite patterns in this study differed from that of dental enamel found on permanent teeth., Competing Interests: The authors declare no conflict of interest., (©2023 The Author(s). Published by MRE Press.)

Exposure of dentin tubules due to loss of protective enamel (crown) and cementum (root) tissues as a result of erosion, mechanical wear, gingival recession, etc. has been the leading causes of dentin hypersensitivity. Despite being a widespread ailment, no permanent solution exists to address this oral condition. Current treatments are designed to alleviate the pain by either using desensitizers or blocking dentin tubules by deposition of minerals or solid precipitates, which often have short-lived effects. Reproducing an integrated mineral layer that occludes exposed dentin with concomitant peritubular mineralization is essential to reestablish the structural and mechanical integrity of the tooth with long-term durability. Here, we describe a biomimetic treatment that promotes dentin repair using a mineralization-directing peptide, sADP5, derived from amelogenin. The occlusion was achieved through a layer-by-layer peptide-guided remineralization process that forms an infiltrating mineral layer on dentin. The structure, composition, and nanomechanical properties of the remineralized dentin were analyzed by cross-sectional scanning electron microscopy imaging, energy dispersive X-ray spectroscopy, and nanomechanical testing. The elemental analysis provided calcium and phosphate compositions that are similar to those in hydroxyapatite. The measured average hardness and reduced elastic modulus values for the mineral layer were significantly higher than those of the demineralized and sound human dentin. The structural integration of the new mineral and underlying dentin was confirmed by thermal aging demonstrating no physical separation. These results suggest that a structurally robust and mechanically durable interface is formed between the interpenetrating mineral layer and underlying dentin that can withstand long-term mechanical and thermal stresses naturally experienced in the oral environment. The peptide-guided remineralization procedure described herein could provide a foundation for the development of highly effective oral care products leading to novel biomimetic treatments for a wide range of demineralization-related ailments and, in particular, offers a potent long-term solution for dentin hypersensitivity.

3D porous hydroxyapatite (HA) has been reinforced by zirconia (ZrO 2 ) coating and impregnation with a combination of platelet rich plasma (PRP) as a source of growth factors (GFs) and Heparin sulfate (HS) to sustain the release of GFs. Adipose mesenchymal stem cells (ADMSCs) were characterized by flow cytometry for CD (cluster of differentiation) 44, CD105, CD106, CD34 and CD144, along with checking the multipotency by differentiation into the adipocytes and osteoblasts. Then, they were cultured on the scaffold treated with and without osteogenic media on days 7, 14 and 21. Electron micrograph and PKH staining show that the ADMSCs have a fusiform phenotype in the absence of osteogenic induction. Cell viability assay shows a higher number of the viable cells on the PRP-containing scaffolds than PRP-free scaffolds on day 7. Colorimetric evaluation, quantitative RT-PCR and immunocytochemistry demonstrate that PRP and HS significantly elevate the alkaline phosphatase enzyme activity and also accelerate the production of both early and mid-osteogenic markers, including collagen I and osteopontin expression with and without osteogenic conditions. The PRP-HS also accelerates the expression of the late osteogenic marker, osteocalcin, in both mRNA and protein level expression with a peak on day 21. In conclusion, supplementation of HA/ZrO 2 with PRP/HS has a synergistic impact on the ADMSCs, even in the absence of chemical induction. It seems that HA/ZrO 2 /PRP/HS scaffold provides a higher osteoconductive microenvironment for stem cell differentiation to osteoblasts., (© 2021. The Author(s), under exclusive licence to Springer Nature B.V.)

The acquired enamel pellicle (AEP) is a multi-protein film attached to the surface of teeth, which functions to lubricate the dental surface, form an anti-erosive barrier and exhibits antimicrobial properties. The initiation of AEP formation occurs within seconds of exposure to saliva, a biofluid rich in protein species. While there have been many publications on the formation of human AEP there is little research on the composition of canine AEP during its acquisition. The aim of these studies was to explore the composition of canine AEP formation, utilising hydroxyapatite (HA) discs as a tooth substitute matrix, over time. Qualitative and quantitative proteomics techniques using tandem mass tag labelled peptides and LC-MS/MS were used to follow the formation of canine AEP on hydroxyapatite discs over the course of an hour. Proteins adsorbed to the HA surface included highly abundant proteins in canine saliva, antimicrobial proteins, protease inhibitors and the buffering agent carbonic anhydrase. Greater understanding of the canine AEP deepens fundamental knowledge of the early processes driving bacterial colonisation of the tooth surface and subsequent plaque accumulation.

Many essential elements exist in nature with significant influence on dentin and bone apatite tissue. Hydroxyapatite (HAp) is the major inorganic crystalline structure of dentin that provides a site for various physiological functions such as surface layer ion exchange. Decades of apatite research have shown that enamel is a high-substituted crystalline apatite, but recent findings suggest that dentin apatite may play a more important role in regulating ion exchange as well as mineral crystallinity. This article is the first part of a review series on the functional role of inorganic trace elements including magnesium, strontium, zinc, and iron in dentin hydroxyapatite. The morphology, physiology, crystallinity, and solubility of these elements as they get substituted into the HAp lattice are extensively discussed. An electronic search was performed on the role of these elements in dentin apatite from January 2007 to September 2021. The relationship between different elements and their role in the mineral upkeep of dentin apatite was evaluated. Several studies recognized the role of these elements in dentinal apatite composition and its subsequent effects on morphology, crystallinity, and solubility. These elements are of great importance in physiological processes and an essential part of living organisms. Magnesium and strontium stimulate osteoblast activity, while zinc can improve overall bone quality with its antibacterial properties. Iron nanoparticles are also vital in promoting bone tissue growth as they donate or accept electrons in redox reactions. Thus, understanding how these elements impact dentin apatite structure is of great clinical significance., (Copyright © 2022 Elsevier GmbH. All rights reserved.)

Although the use of fluoride for root caries control is reported to be effective, the mechanism of maintaining hydroxyapatite is still unclear. This study elucidates the roles of fluoride in the recrystallization of hydroxyapatite, and the impact of calcium to maintain the abundance of hydroxyapatite on acid-challenged root dentin with a novel approach - using synchrotron radiation. Root dentin samples obtained from 40 extracted human premolars were subjected to pH challenge in combination with fluoride treatment. The effect of fluoride on hydroxyapatite regeneration on the root was investigated by using a range of fluoride concentrations (1000-5000 p.p.m.) and the EDTA-chelation technique in vitro. Synchrotron radiation X-ray micro-computed tomography and X-ray absorption spectroscopy were utilized to characterize the chemical composition of calcium species on the surface of prepared samples. The percentage of hydroxyapatite and the relative abundance of calcium species were subsequently compared between groups. The absence of calcium or fluoride prevented the complete remineralization of hydroxyapatite on the surface of early root caries. Different concentrations of fluoride exposure did not affect the relative abundance of hydroxyapatite. Sufficient potency of 1000 p.p.m. fluoride solution in promoting hydroxyapatite structural recrystallization on the root was demonstrated. Both calcium and fluoride ions are prerequisites in a caries-prone environment. Orchestration of F - and Ca 2+ is required for structural homeostasis of root dentin during acid attack. Sustainable levels of F - and Ca 2+ might thus be a strict requirement in the saliva of the population prone to root caries. Fluoride and calcium contribute to structural homeostasis of tooth root, highlighting that routine fluoride use in combination with calcium replenishment is recommended for maintaining dental health. This study also demonstrates that utilization of synchrotron radiation could provide a promising experimental platform for laboratory investigation especially in the dental material research field., (open access.)

This study presents a novel tool to predict temperature-exposure of incinerated pig teeth as a proxy for understanding impacts of fire on human teeth. Previous studies on the estimation of temperature-exposure of skeletal elements have been limited to that of heat-exposed bone. This predictive tool was developed using a multinomial regression model of colourimetric and hydroxyapatite crystal size variables using data obtained from unheated pig teeth and teeth incinerated at 300 °C, 600 °C, 800 °C and 1000 °C. An additional variable based on the observed appearance of the tooth was included in the tool. This enables the tooth to be classified as definitely burnt (600 °C-1000 °C) or uncertain (27 °C/300 °C). As a result, the model predicting the temperature-exposure of the incinerated teeth had an accuracy of 95%. This tool is a holistic, robust and reliable approach to estimate temperature of heat-exposed pig teeth, with high accuracy, and may act as a valuable proxy to estimate heat exposure for human teeth in forensic casework., (© 2021. The Author(s), under exclusive licence to Springer-Verlag GmbH, DE part of Springer Nature.)

Objective: To assess the potential of spectral photon-counting (PC) radiography (SPCR) for the detection and characterization of monosodium urate (MSU) and calcium hydroxyapatite (HA) crystals, based on effective atomic number (Z eff ) values derived from specific X-ray attenuation characteristics at different energy levels., Methods: Suspensions of either pure agar, synthetic MSU (200 mg/ml) or HA (100 and 150 mg/ml) crystals in agar were sealed in industry-standard polystyrene vials and supported on a 2.5-mm-thick plastic table. Samples were scanned using a vendor microfocus X-ray tube and a spectral PC detector prototype with four energy thresholds per acquisition (15, 25, 30, and 35 keV). Material decomposition calibration was performed using polymethyl methacrylate (PMMA) and polyvinylchloride (PVC) slabs. Using a custom post-processing software based on polynomial material decomposition, Z eff of the respective samples were computed. All samples were additionally scanned using dual-energy CT (DECT, 80 kV and tin-filtered 150 kV) and analyzed with a proprietary post-processing algorithm for gout., Results: MSU crystal suspension attenuated significantly less than both HA samples. MSU and HA suspensions differed significantly in Z eff (mean ± SD: 7.74 ± 0.28 vs. 9.43 ± 0.41, p < .001). Z eff values from SPCR were comparable to DECT-based reference values (p = 0.16) and were independent of the radiation dose level (0.18 - 18 mAs, p = 1)., Discussion: This in vitro feasibility study demonstrates the potential of SPCR for discriminating MSU from HA crystal suspensions based on Z eff differences. Further studies have to corroborate these initial findings ex vivo and in vivo, and to compare the diagnostic performance of SPCR with DECT in imaging of crystal-associated arthropathies., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Lead (Pb) complexed with dissolved organic matter (DOM-Pb) is a dominant Pb species in soils, but it is not clear that DOM-Pb is stably immobilized by hydroxyapatite. This study investigates how DOM-Pb is immobilized by hydroxyapatite in both aqueous solutions and soils. A sorption test showed that 69.5% of DOM-Pb is removed in an aqueous solution, but less DOM-Pb is retained by hydroxyapatite compared with Pb ions. On the basis of the ratio of Pb and dissolved organic carbon before and after the sorption test, 7% of Pb was retained as DOM-Pb by hydroxyapatite, but 93% of Pb was dissociated from DOM-Pb, and was sorbed as Pb ions. The concentrations of water-soluble Pb re-released were higher in the DOM-Pb solution than those in the Pb ion solution in sandy loam soil with hydroxyapatite. A column-leaching test that flowed the DOM-Pb solution showed that Pb concentrations in leached water from sandy loam soil gradually increased after the middle stage of the test despite the presence of hydroxyapatite. The amount of water-soluble Pb re-released from soils with and without the hydroxyapatite that flowed the DOM-Pb solution was the same as or greater than that without the hydroxyapatite that flowed the Pb ion solution. This study concludes that in soils with low Pb sorption ability, some of the Pb retained as DOM-Pb is water soluble and possibly re-released despite the presence of hydroxyapatite, although most Pb in DOM-Pb is stably sorbed as Pb ions., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Objectives: Stable isotope studies often focus on hydroxyapatite (bioapatite) to answer questions of paleodiet, paleomobility, and paeloenvironment. This study seeks to determine the effect that sample particle size (in particular SA:V, or surface area to volume ratios) has on measured carbon and oxygen stable isotope values (δ 13 C and δ 18 O) in bone hydroxyapatite., Materials and Methods: Previously ground Homo sapiens sapiens cortical bone samples were subdivided using geological screens to obtain three separate sub-samples, differing only in their particle size. These aliquots (n = 60) were then treated using established protocols to remove any exogenous organic material (2.5% NaOH) and adsorbed carbonates (0.1 M CH 3 COOH), and analyzed for δ 13 C and δ 18 O using a Kiel-IV Carbonate Device coupled to a Thermo-Finnigan DeltaPlus IRMS., Results: Data obtained indicate that decreased particle size leads to increases in both δ 13 C and δ 18 O, with oxygen isotope values being more dramatically affected. Specifically, it is possible to produce isotopic shifts of as much as 1.0‰ and 4.0‰ for δ 13 C and δ 18 O, respectively, solely by analyzing different sized particles from the same individual, bone, and sample., Discussion: Based upon the variability seen between different size fractions from the same sample, it is clear that particle size has a meaningful impact on carbon and oxygen isotope composition. We attribute these shifts to the differential adsorption or precipitation of environmental carbon and oxygen during pretreatment. We recommend that particle size be added to the list of potential variables affecting isotope composition, alongside other factors including diagenesis, reagent concentration, and treatment time. We would also note that while most individuals exhibit consistent changes, some do not, and thus further investigation into these phenomena is warranted., (© 2020 Wiley Periodicals, Inc.)

Fish-scale waste is rich in biocompatible hydroxyapatite (HAp). In the present study, an environmentally friendly method of extracting HAp from fish-scale waste was developed in an effort to promote environmental sustainability. Deep eutectic solvents (choline chloride/glycerol, 1/2) were used to extract HAp from bighead carp (Aristichthys nobilis) scales. A relatively high extraction rate of 47.67% ± 1.81% was obtained under optimum conditions (70 °C, a solid/liquid ratio of 1/15 g/g and a 2.5 hr extraction time). The obtained HAp was characterized and its purity was determined using Fourier transform infrared spectroscopy and X-ray diffraction, respectively. The chemical composition was performed by energy-dispersive X-ray spectrometry and inductively coupled plasma-optical emission spectroscopy. Its morphology and particle size were observed using scanning electron microscopy and particle size distribution analysis. Thermogravimetric analysis was used to determine its thermal stability. Blood compatibility was determined using a hemolytic test. The results showed that this extraction yielded HAp with the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, indicating that the proposed method is an excellent alternative for the improved utilization of fish scale waste. PRACTICAL APPLICATION: Biocompatible hydroxyapatite (HAp) was extracted from fish scale (FS) waste by using an environmentally friendly deep eutectic solvent. The optimized extraction and structure characterization of extracted HAp were investigated in this study. The results showed that the extracted HAp had the irregular morphology, the higher Ca/P ratio, good thermal stability, and blood compatibility, which indicated that the proposed method was an excellent alternative to improving the utilization of FS waste., (© 2019 Institute of Food Technologists®.)

Objectives: To characterize the paleodiet of individuals from Formative Period (1500 B.C.-A.D. 400) Atacama Desert sites of Ancachi and Quillagua as a means of understanding the dietary and cultural impacts of regional systems of exchange., Materials and Methods: Thirty-one bone samples recovered from the cemetery of Ancachi (02QU175) and in/around the nearby town of Quillagua were the subject of carbon and nitrogen stable isotope analysis of bone collagen and hydroxyapatite and multisource mixture modeling (FRUITS, food reconstruction using isotopic transferred signals) of paleodiet. These individuals were compared with nearly 200 other Formative Period individuals from throughout the region to identify differences in dietary behaviors., Results: 80.6% (25/31) of the samples yielded sufficient well-preserved collagen and were included in the multisource mixture model. The FRUITS model, which compared individuals with a robust database of available foods from the region, identified a wide diversity of diets in the Ancachi/Quillagua area (including both coastal and interior individuals), and, most notably, thirteen individuals who consumed an average of 11.2 ± 1.9% terrestrial animals, 19.8 ± 1.9% legumes, and 22.5 ± 3.1% marine fauna, a balanced pattern of protein consumption distinct from both the coastal and inland individuals in our larger regional sample., Conclusions: The combination of stable isotope analysis and multisource mixture modeling permitted the characterization of dietary behavior of 25 individuals from nodal sites in the Atacama Desert, thus enhancing our understanding of the economic and social relationships that bound together Formative Period sites, populations, and individuals in this hyperarid region., (© 2019 Wiley Periodicals, Inc.)

Osteoarthritis (OA) is not only related to the degradation of articular cartilage, but also possibly to the changes of subchondral bone. The purpose of this study was to assess whether specific differences could be resolved from bone composition, as also contributed to OA. These differences were assessed by using Fourier transform infrared spectroscopy (FTIRS). The main parameters including mineral content, carbonate content, crystallinity, collagen cross-linking ratio (XLR) and acid phosphate content were represented with characteristic peak integration. It was found that mineral and carbonate content varied significantly with depths at different OA stages. Mineral content increased with depth in healthy samples, while carbonate content showed opposite trend. The mineral content reduced obviously with OA duration, which was different with carbonate decreasing only at early stage of OA. In addition, the content of acid phosphate, collagen maturity (XLR) and crystallinity slight varied with the OA aggravation. Therefore, the changes in subchondral bone were significantly associated with cartilage degeneration and OA, the associated parameters should be targeted for OA therapies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

The purpose of this study was to evaluate the micro-tensile bond strength (μTBS) of dentin bonding agents containing different ratios of nano-sized hydroxyapatite fillers (HA). X-ray diffraction analysis was used for characterization, and scanning electron microscope (SEM) analysis was used to determine the HA particle size after that HA were mixed a bonding agents without filler. Dentin bonding agents were divided into four groups according to addition of different ratios of nano-sized hydroxyapatite fillers as 2% HA, 5% HA, 7% HA, and no-filler control group. The teeth (n = 32) were sectioned with a low-speed diamond blade under water cooling to expose the mid-coronal dentin. Following the bonding application, restorations were applied incrementally. Each tooth was cut on the x and y axis, and each specimen was fixed to a testing device and stressed until failure occurred. The debonded specimens were examined under 250× magnification without a coating layer at 2.00 kV using a SEM to determine failure patterns. μTBS data were analyzed using a anova and Tukey's post hoc test. The failure mode data were analyzed using the Chi-Square test. The maximum mean value of μTBS was in the 7% HA group, while the minimum mean value of μTBS was observed in the control group. 7% HA group was statistically significant and higher than other groups while there were no significant differences between the control, 2% HA, and 5% HA groups. According to SEM analysis, fracture analysis revealed that the mixed fracture type was seen more often than the other fracture types. The particle size and amount of HA fillers added to the adhesive resin seem to affect the success of the bond strength to the dentin. Adding different ratio nano-sized HA fillers to the adhesive resin contributed positively to the immediate μTBS values in the dentin., (© 2019 Wiley Periodicals, Inc.)

Goal: Evaluate bone regeneration in a critical size bone defect model in the jaw of healthy rats as a function of gender and defect location., Design: A series of microCT and histological studies were performed to evaluate the process of bone regeneration in rats with a mandibular critical size defect. Rats were placed in two groups according to gender and sorted in terms of bone defect location. Bone regeneration rate and hydroxyapatite concentration were assessed with microCT imaging at specific times after surgery. Histological analysis was also performed to evaluate bone regeneration., Results: No more that 85% of bone regeneration was observed after 60 days, with a low rate constant (K) indicating a slow restoration of the defect. Assessment of microCT images showed partial closure of the defect in all cases, which was confirmed by histological analysis. Hydroxyapatite concentration values revealed that regenerated bone was not fully calcified. No statistically significant differences in terms of gender or defect location were found., Conclusion: The defect model studied here, located in the jaw of healthy rats, shows potential as a preclinical critical size bone defect model to evaluate bone regeneration therapies in the fields of dentistry and maxillofacial surgery., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Vibrational micro-spectroscopy is a powerful optical tool, providing a non-invasive label-free chemically specific imaging for many chemical and biomedical applications. However, hyperspectral image produced by Raman micro-spectroscopy typically consists of thousands discrete pixel points, each having individual Raman spectrum at thousand wavenumbers, and therefore requires appropriate image unmixing computational methods to retrieve non-negative spatial concentration and corresponding non-negative spectra of the image biochemical constituents. Here, we present a new efficient Quantitative Hyperspectral Image Unmixing (Q-HIU) method for large-scale Raman micro-spectroscopy data analysis. This method enables to simultaneously analyse multi-set Raman hyperspectral images in three steps: (i) Singular Value Decomposition with innovative Automatic Divisive Correlation which autonomously filters spatially and spectrally uncorrelated noise from data; (ii) a robust subtraction of fluorescent background from the data using a newly developed algorithm called Bottom Gaussian Fitting; (iii) an efficient Quantitative Unsupervised/Partially Supervised Non-negative Matrix Factorization method, which rigorously retrieves non-negative spatial concentration maps and spectral profiles of the samples' biochemical constituents with no a priori information or when one or several samples' constituents are known. As compared with state-of-the-art methods, our approach allows to achieve significantly more accurate results and efficient quantification with several orders of magnitude shorter computational time as verified on both artificial and real experimental data. We apply Q-HIU to the analysis of large-scale Raman hyperspectral images of human atherosclerotic aortic tissues and our results show a proof-of-principle for the proposed method to retrieve and quantify the biochemical composition of the tissues, consisting of both high and low concentrated compounds. Along with the established hallmarks of atherosclerosis including cholesterol/cholesterol ester, triglyceride and calcium hydroxyapatite crystals, our Q-HIU allowed to identify the significant accumulations of oxidatively modified lipids co-localizing with the atherosclerotic plaque lesions in the aortic tissues, possibly reflecting the persistent presence of inflammation and oxidative damage in these regions, which are in turn able to promote the disease pathology. For minor chemical components in the diseased tissues, our Q-HIU was able to detect the signatures of calcium hydroxyapatite and β-carotene with relative mean Raman concentrations as low as 0.09% and 0.04% from the original Raman intensity matrix with noise and fluorescent background contributions of 3% and 94%, respectively., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Osteosarcoma is the most common primary malignant bone tumor. In the last years, several studies have demonstrated that the increase of Hydroxyapatite (HA) and Interleukin-6 (IL-6) syntheses compared to those expressed by normal osteoblasts could be used to detect the degree of malignancy of osteosarcoma cells. Conventional biochemical methods widely employed to evaluate bone cell differentiation, including normal and cancerous phenotypes, are time consuming and may require a large amount of cells. HA is a mineral form of calcium phosphate whose presence increases with maturation of osteosarcoma cells. Analogously, IL-6 is a fundamental cytokine whose production is highly increased in osteosarcoma cells. In this study, we employ Raman spectroscopy to the identification and discrimination of osteosarcoma cells from osteo-differentiated mesenchymal stromal cells (MSCs) by detecting the presence of HA and IL-6. However, while the identification of HA is facilitated by the characteristic peak at 960 cm -1 , corresponding to symmetric stretching (P-O) mode, the quantification of IL-6 it is much more elusive, being its Raman signal characterized by cysteine, but also by phenylalanine, amide I II and III whose signals are common to other proteins. Supported by an accurate multivariate analysis, the results show that Raman spectroscopy is a high sensitivity technique dealing out a direct and quantitative measurement of specific mineralization levels of osteosarcoma cells. In turn, by exploiting the Surface-Enhanced Raman Scattering stimulated by internalized Gold Nanoshells (AuNSs) and combined with scanning probe microscopies, we were able to employ Raman spectroscopy to study subcellular components locally.

Thermal neutron radiography and X-ray radiography are characterised by different penetration depths in various materials, for example in collagen and hydroxyapatite, two major components of bone. Neutron radiography penetrates hydroxyapatite easier than collagen and, conversely, in X-ray radiography attenuation is higher in hydroxyapatite than in collagen. This effect allows estimation of collagen presence in dry bone. In our study we show that differences between the two imaging methods are sufficient to produce significant results when bone areas with higher and lower content of collagen are being compared., (Copyright © 2018 Elsevier Ltd. All rights reserved.)

Hydroxyapatite (HA) material will be long-standing once implanted in bone tissue of the body. It should be considered to endow the osteogenic HA material with traceable fluorescence to realize a lifelong in vivo tracking. We prepared and utilized lanthanides-doped HA upconversion material, and revealed for the first time that the lanthanides (ytterbium (Yb) and holmium (Ho)) co-doped HA upconversion material was suitable for long-term or lifelong in vivo tracking, the lanthanide ions doped in the HA matrix would not affect the biocompatibility and osteogenesis, and the tissue autofluorescence could be effectively avoided by the HA:Yb/Ho upconversion material. Also the distribution in bone and osteointegration with bone of the HA:Yb/Ho material could be clearly discriminated by its bright fluorescence under NIR irradiation. The upconversion characteristic of the HA:Yb/Ho material provides a feasibility and promising prospect for lifelong in vivo tracking, and has an advantage in revealing the material-tissue interrelation. The material has important clinical application value in addition to its usefulness for scientific investigation.

Whether hydroxyapatite (HA) orientation in fossilised bone samples can be non-destructively retrieved and used to determine the arrangement of the bone matrix and the location of muscle attachments (entheses), is a question of high relevance to palaeontology, as it facilitates a detailed understanding of the (micro-)anatomy of extinct species with no damage to the precious fossil specimens. Here, we report studies of two fossil bone samples, specifically the tibia of a 300-million-year-old tetrapod, Discosauriscus austriacus, and the humerus of a 370-million-year-old lobe-finned fish, Eusthenopteron foordi, using XRD-CT - a combination of X-ray diffraction (XRD) and computed tomography (CT). Reconstructed 3D images showing the spatial mineral distributions and the local orientation of HA were obtained. For Discosauriscus austriacus, details of the muscle attachments could be discerned. For Eusthenopteron foordi, the gross details of the preferred orientation of HA were deduced using three tomographic datasets obtained with orthogonally oriented rotation axes. For both samples, the HA in the bone matrix exhibited preferred orientation, with the unit cell c-axis of the HA crystallites tending to be parallel with the bone surface. In summary, we have demonstrated that XRD-CT combined with an intuitive reconstruction procedure is becoming a powerful tool for studying palaeontological samples.

Purpose: The paper focuses on the relationship of risk factors and metabolic disorders with mineralogical composition of calculi, age and gender of calcium oxalate stone formers., Materials and Methods: Stone mineralogical composition, 24 hour biochemistry and pH-profile of urine were examined for sixty four stone formers using powder X-ray diffraction, spectrophotometric and potentiometric techniques., Results: The analysis indicated that 44 % of calculi were composed of pure calcium oxalate monohydrate, whereas other 56 % contained both monohydrate and dihydrate or usually their mixtures with hydroxyl apatite. Hypocitraturia, hypercalciuria and hyperuricosuria were identified as the most frequent disorders. Patients with pure calcium oxalate stones and calcium oxalate mixed with apatite revealed different patterns including age, acid-base balance of urine, calcium, citrate excretion etc.Conclusions: Our results demonstrate that most patients simultaneously reveal several risk factors. The special attention should be paid to normalize the daily citrate, calcium and urate excretion. High risk patients, such as postmenopausal females or stone formers with a high apatite content require a specific metabolic evaluation towards in highlighting abnormalities associated with stone formation.

Background: Many investigators reported the amount of fluoride release from glass ionomer cement. However, the work on fluoride release from GIC containing fluoroapatite and hydroxyapatite is scarce. Therefore, this study was conducted to find out the amount of fluoride release from Glass ionomer cement containing fluoroapatite and hydroxyapatite., Methods: The study was conducted in the Department of Materials, Queen Marry University of London. A total of 108 samples equally divided in to three groups namely fluoroapatite added GIC, Hydroxyapatite added GIC as an experimental group and unmodified GIC as a control group. The specimens were prepared by mixing powder and liquid in the ratio of 1:1. Amount of fluoride released was measured by Ion electrode method at 1, 3, 7, 14, 21 and 28 days., Results: On day 1, the combination of FA +GIC showed the highest amount of fluoride release followed by the control group (GIC) whereas the combination of HA+GIC released the least amount of fluoride. On day 7, the amount of fluoride release started declining in all three groups. The amount of fluoride release continued decreasing on day 21 in which combination of FA +GIC and the control group are shown to release equal amount of fluoride whereas the combination of HA+GIC gave the least activity the amount of fluoride release fall to a minimum level in all three group by day 28., Conclusions: It is concluded that addition of fluoroapatite into GIC has significant effect on the amount of fluoride release as compared to GIC alone; however, addition of hydroxyapatite into GIC has no additive effect on the amount of fluoride release.

Aim: To characterize chemical degradation of the principal constituents of dentine after exposure to NaOCl and EDTA using Infrared Spectroscopy (ATR-FTIR)., Methodology: Ground dentine particles, from extracted permanent human molars, were passed through sieves of 38 to 1 000 μm to provide six size ranges. Portions (250 mg) of each size range were reacted with 5 mL of 2.5% NaOCl for 2-10 min; or 17% EDTA for 5-1440 min. Powders larger than 75 μm were also sequentially exposed to NaOCl/EDTA/NaOCl each for 10 min. All experiments were repeated five times. Reacted and unreacted powders were washed and dried. Particles larger than 75 μm were then reground. FTIR spectra of unground and reground reacted particles enabled assessment of particle surface versus bulk chemistry, respectively, plus estimation of reaction depth. Changes in the ratio of the 1 640 cm -1 collagen: 1 010 cm -1 phosphate peak height or its inverse were obtained. These were used to estimate surface and bulk fraction reacted and thus depth to which collagen or phosphate was reduced following immersion in NaOCl or EDTA, respectively. The data were analysed descriptively., Results: Surface collagen fraction declined by ~40% within 2 min of NaOCl exposure, and plateaued at ~60% between 6-10 min. Bulk spectra showed average depth of collagen loss at 10 min was 16 ± 13 μm. Ten minute EDTA exposure caused ~60% loss of surface phosphate. Average depth of phosphate loss was 19 ± 12 μm and 89 ± 43 μm after 10 and 1 440 min EDTA immersion, respectively. Sequential NaOCl/EDTA immersion yielded a 62 ± 28 μm thick phosphate-depleted surface. Sequential NaOCl/EDTA/NaOCl treatment resulted in approximately 85 μm of collagen loss., Conclusions: Data revealed the sequential depletion of collagen by NaOCl and apatite by EDTA in dentine, simultaneously exposing the other moieties. Alternate exposure to NaOCl and EDTA therefore enhances the depth of erosion., (© 2017 International Endodontic Journal. Published by John Wiley & Sons Ltd.)

Background: Microcalcifications within the fibrous cap of the arteriosclerotic plaques lead to the accrual of plaque-destabilizing mechanical stress. New techniques for plaque screening with small detectors and the ability to differentiate between the smooth and hard elements of plaque formation are necessary., Method: Vascular plaque formations are characterized as calcium phosphate containing structures organized as hydroxylapatite resembling the mineral whitlockite. In transmission and reflexion studies with a simple millimeter wave (mm-wave)-demonstrator, we found that there is a narrow window for plaque detection in arterial vessels because of the tissue water content, the differentiation to fatty tissue, and the dielectric property of air or water, respectively., Result: The new sensor is based on a sensing oscillator working around 27 GHz. The open-stub capacitance determines the operating frequency of the sensor oscillator. The capacitance depends on the dielectric properties of the surrounding material. The sensor components were completely built up in surface mount technique., Conclusion: Completed with a catheter, the sensor based on microwave technology appears as a robust tool ready for further clinical use., Competing Interests: Conflict of Interest: None declared., (Georg Thieme Verlag KG Stuttgart · New York.)

The purpose of this study is to analyze the mechanical enhancement provided by nanocomposite coatings deposited on robocast 45S5 bioglass (BG) scaffolds for bone tissue regeneration. In particular, a nanocomposite layer consisting of hydroxyapatite (HA) nanoparticles, as reinforcing phase, in a polycaprolactone (PCL) matrix was deposited onto the surface of the BG struts conforming the scaffold. Three different HA nanopowders were used in this study. The effect of particle size and morphology of these HA nanopowders on the mechanical performance of 45S5 BG scaffolds is evaluated., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Purpose: The goal of this study was to compare the in vitro bioactivity in simulated body fluid (SBF) of commercially available dental implants., Materials and Methods: Bioactivity, according to ISO 23317, of commercially available dental implants with various surface modifications (BIO-surface, SLA, SLActive, TiUnite, and OsseoSpeed) was tested in SBF for 1 and 3 weeks. Surface characterizations, especially calcium and phosphorus surface content before and after the immersion in SBF, were performed. The effect of surface treatment on bioactivity was studied., Results: Differences between surfaces before immersion in SBF were confirmed by Raman spectroscopy, x-ray photoelectron spectroscopy (XPS), energy-dispersive x-ray spectroscopy (EDX), and scanning electron microscope (SEM) analysis. Calcium and phosphorus surface content was increasing within the tested period in the case of two (BIO-surface and SLActive) of the five tested dental implants. Calcium-phosphate precipitation was observed by SEM, XPS, EDX, and x-ray micro‑diffraction (μ-XRD) analysis., Conclusion: Two (BIO-surface from LASAK and SLActive from Straumann) of the five tested dental implants were found to be bioactive, according to ISO 23317. Although it is difficult to unambiguously determine the properties that have influence on the hydroxyapatite precipitation rate, multiple properties that the two surfaces have in common were found.

Ti-Nb-Sn-hydroxyapatite (HA) composites were prepared by mechanical alloying for different times (unmilled, 4, 8 and 12h), followed by pulse current activated sintering. The effects of the milling time on the electrochemical corrosion resistance and bioactivity of the sintered Ti-35Nb-2.5Sn-15HA composites were investigated. Potentiodynamic polarization test results indicated that the sintered Ti-35Nb-2.5Sn-15HA composites exhibited higher corrosion resistance with increasing milling time. The corrosion potential and current of the Ti-35Nb-2.5Sn-15HA composite sintered by 12h milled powders were - 0.261V and 0.18μA/cm 2 , respectively, and this sintered composite showed a stable and wide passivation region. The hemolysis rate of the sintered Ti-35Nb-2.5Sn-15HA composites reduced with increasing milling time and the lowest hemolytic rate of the composites was 0.87%. In addition, the in vitro cell culture results indicated that the composite sintered by 12h milled powders had good biocompatibility. These results indicate the significant potential of Ti-35Nb-2.5Sn/xHA composites for biomedical implant applications., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Archival research into episodes of frontier violence in the Kimberley region of Western Australia indicate that the bodies of Aboriginal victims of massacres were frequently incinerated following the event. This paper presents the results of a scientific investigation of a reported massacre at Sturt Creek where burnt bone fragments were identified in two adjacent sites and documents the archaeological signatures associated with the sites. The methodology used to undertake the project brought together three systems of knowledge: the oral testimonies of the descent group originating from a sole adult survivor of the massacre; archival, historical and scientific research. An archaeological survey defined the two distinct sites containing hundreds of fragile bone fragments; a third site was found to be highly disturbed. Scientific investigations included macroscopic and microscopic examination of selected bone fragments by an anatomical pathologist and a zooarchaeologist and X-ray diffraction analysis of sixteen bone fragments. The anatomical pathologist and zooarchaeologist undertook macroscopic and microscopic examinations of selected bone samples to identify morphological evidence for human origin. It was concluded that three bone fragments examined may have been human, and two of the fragments may have been from the vault of a skull. It was concluded that the likelihood of them being human would be strengthened if it was found that the three samples had been subjected to high temperatures. X-ray diffraction analysis of 16 bone fragments provided this evidence. All fragments showed sharp hydroxylapatite peaks (crystallite sizes 9882nm and 597nm respectively) and all had been subjected to extreme temperatures of either 600°C for more than 80h, 650°C for more than 20h, 700°C for more than 4h or 800°C for more than 1h. XRD analyses were also done on bone samples collected from three cooking hearths at three different archaeological sites. It was found that two of the three samples had been exposed to substantially lower temperatures for a short time period. It was concluded that there was strong pathological and archaeological evidence that the bone fragments were human in origin, but that the evidence was not conclusive. This research also identified archaeological signatures for the identification of massacre sites in similar Australian environments and circumstances., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Coronary calcification is a risk factor for ischemic heart disease. Hydroxyapatite that is formed by polymerization from calcium phosphate tribasic (CPT) is the major constituent of coronary calcium deposits. If CPT could be visualized, coronary calcification could be predicted and prevented. We discovered that when CPT and collagen I, the main constituent of collagen fibers, are mixed with lac dye (LD) and then exposed to fluorescent light excited at 345 ± 15 nm and emitted at 420 nm, a purple fluorescence that is characteristic of CPT only is elicited. So, we examined localization of CPT and its relation to plaque morphology by color fluorescent angioscopy (CFA) or microscopy (CFM) in 24 coronary arteries obtained from 12 autopsy subjects. By CFA, the incidence (%) of CPT as confirmed by purple fluorescence in 15 normal segments, 25 white plaques, 14 yellow plaques without necrotic core (NC) and 8 yellow plaques with NC was 20, 36, 64 and 100 (p < 0.05 vs. normal segments), respectively. By CFM, the CPT was either deposited alone amorphously or surrounded hydroxyapatite that was identified by Oil Red O, methylene blue and von Kossa's stain. The results suggested that CFA using LD is feasible for imaging CPT, that is a precursor of hydroxyapatite, in human coronary plaques, and this technique would help prediction and discovery of a preventive method of coronary calcification.

Raman and Fourier transform infrared (FT-IR) spectroscopic imaging techniques can be used to characterize bone composition. In this study, our objective was to validate the Raman mineral:matrix ratios (ν 1 PO 4 :amide III, ν 1 PO 4 :amide I, ν 1 PO 4 :Proline + hydroxyproline, ν 1 PO 4 :Phenylalanine, ν 1 PO 4 :δ CH 2 peak area ratios) by correlating them to ash fraction and the IR mineral:matrix ratio (ν 3 PO 4 :amide I peak area ratio) in chemical standards and native bone tissue. Chemical standards consisting of varying ratios of synthetic hydroxyapatite (HA) and collagen, as well as bone tissue from humans, sheep, and mice, were characterized with confocal Raman spectroscopy and FT-IR spectroscopy and gravimetric analysis. Raman and IR mineral:matrix ratio values from chemical standards increased reciprocally with ash fraction (Raman ν 1 PO 4 /Amide III: P < 0.01, R 2  = 0.966; Raman ν 1 PO 4 /Amide I: P < 0.01, R 2  = 0.919; Raman ν 1 PO 4 /Proline + Hydroxyproline: P < 0.01, R 2  = 0.976; Raman ν 1 PO 4 /Phenylalanine: P < 0.01, R 2  = 0.911; Raman ν 1 PO 4 /δ CH 2 : P < 0.01, R 2  = 0.894; IR P < 0.01, R 2  = 0.91). Fourier transform infrared mineral:matrix ratio values from native bone tissue were also similar to theoretical mineral:matrix ratio values for a given ash fraction. Raman and IR mineral:matrix ratio values were strongly correlated ( P < 0.01, R 2  = 0.82). These results were confirmed by calculating the mineral:matrix ratio for theoretical IR spectra, developed by applying the Beer-Lambert law to calculate the relative extinction coefficients of HA and collagen over the same range of wavenumbers (800-1800 cm -1 ). The results confirm that the Raman mineral:matrix bone composition parameter correlates strongly to ash fraction and to its IR counterpart. Finally, the mineral:matrix ratio values of the native bone tissue are similar to those of both chemical standards and theoretical values, confirming the biological relevance of the chemical standards and the characterization techniques.

Background and Aims: Recent studies highlighted the role of calcification processes in the clinical progression of chronic cardiovascular diseases. In this study we investigated the relationship between the chemical composition of calcification and atherosclerotic plaque stability in carotid arteries., Methods and Results: To this end, we characterized the calcification on 229 carotid plaques, by morphology, immunohistochemistry, transmission electron microscopy and energy dispersive X-ray microanalysis. Plaques were classified into two categories: unstable and stable. No significant differences were found in the incidence of the various risk factors between patients with and without carotid calcification, with the exception of diabetes. The energy dispersive X-ray microanalysis allowed us to identify two types of calcium salts in the atheromatous plaques, hydroxyapatite (HA) and calcium oxalate (CO). Our results showed that calcification is a common finding in carotid plaques, being present in 77.3% of cases, and the amount of calcium is not a factor of vulnerability. Noteworthy, we observed an association between HA calcification and unstable plaques. On the contrary, CO calcifications were mainly detected in stable plaques., Conclusions: The presence of different types of calcification in atheromatous plaques may open new perspectives in understanding the molecular mechanisms of atheroma formation and plaque instability., (Copyright © 2017 The Italian Society of Diabetology, the Italian Society for the Study of Atherosclerosis, the Italian Society of Human Nutrition, and the Department of Clinical Medicine and Surgery, Federico II University. Published by Elsevier B.V. All rights reserved.)

Developing a drug carrier system which could perform targeted and controlled release over a period of time is utmost concern in the pharmaceutical industry. This is more relevant when designing drug carriers for poorly water soluble drug molecules such as curcumin and 6-gingerol. Development of a drug carrier system which could overcome these limitations and perform controlled and targeted drug delivery is beneficial. This study describes a promising approach for the design of novel pH sensitive sodium alginate, hydroxyapatite bilayer coated iron oxide nanoparticle composite (IONP/HAp-NaAlg) via the co-precipitation approach. This system consists of a magnetic core for targeting and a NaAlg/HAp coating on the surface to accommodate the drug molecules. The nanocomposite was characterized using FT-IR spectroscopy, X-ray diffraction, scanning electron microscopy, transmission electron microscopy and thermogravimetric analysis. The loading efficiency and loading capacity of curcumin and 6-gingerol were examined. In vitro drug releasing behavior of curcumin and 6-gingerol was studied at pH 7.4 and pH 5.3 over a period of seven days at 37°C. The mechanism of drug release from the nanocomposite of each situation was studied using kinetic models and the results implied that, the release is typically via diffusion and a higher release was observed at pH 5.3. This bilayer coated system can be recognized as a potential drug delivery system for the purpose of curcumin and 6-gingerol release in targeted and controlled manner to treat diseases such as cancer., (Copyright © 2017 Elsevier B.V. All rights reserved.)

To obtain hydroxyapatite (HA) coatings with high crystallinity which have long-term stability in clinical applications, coarse powders were usually injected to less energetic plasma. However, the HA coatings accumulated by partly melted particles usually have high porosity and poor mechanical properties, especially poor bonding strength. In this work, by profiting its quenching and mechanical impact, dry-ice blasting was in-situ employed during plasma spray process to improve the microstructure characterization and bonding strength of HA coatings. In addition, the influence of in-situ dry-ice blasting on the phase composition and crystallinity of plasma-sprayed HA coatings was investigated. The results show that a significant reduction of porosity and an apparent increase in bonding strength are revealed in plasma-sprayed HA coatings due to the cleaning effect of dry-ice blasting on the convex unmelted particles and splashing fragments. HA coatings prepared by the combination process of plasma spraying and dry-ice blasting have a compromise structure with minimum globular pores but with pronounced microcracks. The disappearance of CaO phase and the increase in crystallinity also derive from the application of dry-ice blasting., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In the present paper, first results of the influence of the degradation of biodegradable materials on the hardness of the bone are presented in detail. For this purpose, different materials (Mg, Ti and biopolymers) were implanted into the femora of growing rats and bone cross sections were examined for the micro-hardness (MH). The aim of the present paper was to examine the mechanical response of the bone areas surrounding the implant at defined sites and at specified periods after implantation. A special focus was set on Mg alloys. In earlier in-vitro and in-vivo studies, an accumulation of Magnesium in the vicinity of the implant was detected by using different techniques. Therefore, micro-hardness measurements were performed, and the mechanical strength of bone was correlated with the exchange of Magnesium and Calcium in Hydroxyapatite. After the operation and implantation, the micro-hardness values became temporarily lower, but after complete degradation of the implants, the values were identical with those of specimens containing no implants., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Poly-D-L-lactide/nano-hydroxyapatite (PDLLA/nano-HA) can be used as the biological scaffold material in bone tissue engineering as it can be readily made into a porous composite material with excellent performance. However, constitutive modeling for the mechanical response of porous PDLLA/nano-HA under various stress conditions has been very limited so far. In this work, four types of fundamental compressible hyper-elastic constitutive models were introduced for constitutive modeling and investigation of mechanical behaviors of porous PDLLA/nano-HA. Moreover, the unitary expressions of Cauchy stress tensor have been derived for the PDLLA/nano-HA under uniaxial compression (or stretch), biaxial compression (or stretch), pure shear and simple shear load by using the theory of continuum mechanics. The theoretical results determined from the approach based on the Ogden compressible hyper-elastic constitutive model were in good agreement with the experimental data from the uniaxial compression tests. Furthermore, this approach can also be used to predict the mechanical behaviors of the porous PDLLA/nano-HA material under the biaxial compression (or stretch), pure shear and simple shear., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Titanium (Ti)-hydroxyapatite (HA) composites have the potential for orthopedic applications due to their favorable mechanical properties, excellent biocompatibility and bioactivity. In this work, the pure Ti and nano-scale HA (Ti-nHA) composites were in-situ prepared by selective laser melting (SLM) for the first time. The phase, microstructure, surface characteristic and mechanical properties of the SLM-processed Ti-nHA composites were studied by X-ray diffraction, transmission electron microscope, atomic force microscope and tensile tests, respectively. Results show that SLM is a suitable method for fabricating the Ti-nHA composites with refined microstructure, low modulus and high strength. A novel microstructure evolution can be illustrated as: Relatively long lath-shaped grains of pure Ti evolved into short acicular-shaped and quasi-continuous circle-shaped grains with the varying contents of nHA. The elastic modulus of the Ti-nHA composites is 3.7% higher than that of pure Ti due to the effect of grain refinement. With the addition of 2% nHA, the ultimate tensile strength significantly reduces to 289MPa but still meets the application requirement of bone implants. The Ti-nHA composites exhibit a remarkable improvement of microhardness from 336.2 to 600.8 HV and nanohardness from 5.6 to 8.3GPa, compared to those of pure Ti. Moreover, the microstructure and property evolution mechanisms of the composites with the addition of HA were discussed and analyzed. It provides some new knowledge to the design and fabrication of biomedical material composites for bone implant applications., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

The human head can be subjected to numerous impact loadings such as those produced by a fall or during sport activities. These accidents can result in skull fracture and in some complex cases, part of the skull may need to be replaced by a biomedical implant. Even when the skull is not damaged, such accidents can result in brain swelling treated by decompressive craniectomy. Usually, after recovery, the part of the skull that has been removed is replaced by a prosthesis. In such situations, a computational tool able to analyse the choice of prosthesis material depending on the patient's specific activity has the potential to be extremely useful for clinicians. The work proposed here focusses on the development and use of a numerical model for the analysis of cranial implants under impact conditions. In particular, two main biomaterials commonly employed for this kind of prosthesis are polyether-ether-ketone (PEEK) and macroporous hydroxyapatite (HA). In order to study the suitability of these implants, a finite element head model comprising scalp, skull, cerebral falx, cerebrospinal fluid and brain tissues, with a cranial implant replacing part of the skull has been developed from magnetic resonance imaging data. The human tissues and these two biocompatible materials have been independently studied and their constitutive models are provided here. A computational model of the human head under impact loading is then implemented and validated, and a numerical comparison of the mechanical impact response of PEEK and HA implants is presented. This comparison was carried out in terms of the effectiveness of both implants in ensuring structural integrity and preventing traumatic brain injury. The results obtained in this work highlight the need to take into account environmental mechanical considerations to select the optimal implant depending on the specific patient: whereas HA implants present attractive biointegration properties, PEEK implant can potentially be a much more appropriate choice in a demanding mechanical life style. Finally, a novel methodology is proposed to assess the need for further clinical evaluation in case of impact with both implants over a large range of impact conditions., (Copyright © 2017 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Magnesium/hydroxyapatite composites were produced by conventional extrusion and their mechanical behavior studied under uniaxial compression at room temperature. The results evidence the capability of the HA for strengthening the Mg material, lowering its microstructural anisotropy and inhibiting deformation twinning. They also reveal that the ECAP processing is effective for improving the grain structure and reducing the crystallographic texture of these composites, giving rise to a significant enhancement of their yield strength and microhardness although the ultimate compressive stress worsens. The analysis of the strain hardening rate of the flow curves demonstrates that the HA addition and the ECAP processing are also effective in inhibiting non-basal dislocation slip., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Objectives: Calcium hydroxyapatite (HA) is reported to be the major constituent of soft tissue calcinosis in patients with scleroderma (systemic sclerosis, SSc). Mechanical stress and local tissue hypoxia are thought to be important in the pathogenesis of SSc calcinosis. We sought to analyse spontaneously draining material from calcinosis sites in SSc patients using X-ray diffraction (XRD)., Method: With approval from our local Institutional Review Board (IRB), eligible SSc patients consented to submit their spontaneously draining calcinosis samples for XRD analysis. All patients met the American College of Rheumatology (ACR) criteria for definite SSc. XRD data were used to determine how much solid phase was present (e.g. HA vs. other calcium phosphate phases) and the percentage of crystalline components. Pertinent clinical data were collected., Results: Draining calcinosis was submitted mostly from the hands of 10 female patients with advanced SSc, whose mean disease duration was 16 (range 9-29) years; six had diffuse cutaneous SSc and four had limited SSc. All 10 developed calcinosis later in their disease course; seven had extensive deposits affecting multiple sites. XRD confirmed that HA was the only crystalline material. However, HA was the minor component and most samples contained more than 50% organic material. Solid samples generally contained more HA and fluid samples contained HA in suspension., Conclusions: In this large series of SSc calcinosis analysis, XRD confirmed that HA was the only inorganic material that formed less than 50% of most draining samples. More research is needed to fully characterize and improve our understanding of calcinosis formation in SSc.

The fast, high-throughput distinction between palaeoanthropological/archaeological remains and recent forensic/clinical bone samples is of vital importance in the field of medico-legal science. In this paper, a novel dating method was developed using the autofluorescence of human bones and the confocal laser scanning microscope as the means to distinguish between archaeological and forensic anthropological skeletal findings. Human bones exhibit fluorescence, typically induced by natural antibiotics that are absorbed by collagen, and provide secondary, exogenous fluorophores. However, primary natural fluorescence (or autofluorescence) caused by enigmatic endogenous fluorophores is also present as a micro-phenomenon, whose nature is still obscure. Here, we show that the endogenous fluorophores are mucopolysaccharides of the Rouget-Neumann sheath and, more relevant, that the intensity of the natural fluorescence in human bone decreases in a relationship to the antiquity of the samples. These results suggest that the autofluorescence of bone is a promising technique for the assessment of skeletal remains that may be potentially of medico-legal interest. A larger study is proposed to confirm these findings and to create a predictive model between the autofluorescence intensity and the time since death., (Copyright © 2017 Elsevier Ireland Ltd. All rights reserved.)

We investigated the feasibility of using spatially offset Raman spectroscopy (SORS) for nondestructive characterization of bone tissue engineering scaffolds. The deep regions of these scaffolds, or scaffolds implanted subcutaneously in live animals, are typically difficult to measure by confocal Raman spectroscopy techniques because of the limited depth penetration of light caused by the high level of light scattering. Layered samples consisting of bioactive glass foams (IEIC16), three-dimensional (3D)-printed biodegradable poly(lactic-co-glycolic acid) scaffolds (PLGA), and hydroxyapatite powder (HA) were used to mimic nondestructive detection of biomineralization for intact real-size 3D tissue engineering constructs. SORS spectra were measured with a new SORS instrument using a digital micromirror device (DMD) to allow software selection of the spatial offsets. The results show that HA can be reliably detected at depths of 0-2.3 mm, which corresponds to the maximum accessible spatial offset of the current instrument. The intensity ratio of Raman bands associated with the scaffolds and HA with the spatial offset depended on the depth at which HA was located. Furthermore, we show the feasibility for in vivo monitoring mineralization of scaffold implanted subcutaneously by demonstrating the ability to measure transcutaneously Raman signals of the scaffolds and HA (fresh chicken skin used as a top layer). The ability to measure spectral depth profiles at high speed (5 s acquisition time) and the ease of implementation make SORS a promising approach for noninvasive characterization of cell/tissue development in vitro, and for long-term in vivo monitoring the mineralization in 3D scaffolds subcutaneously implanted in small animals.

The mechanical behavior of bone is determined at all hierarchical levels, including lamellae (the basic building block of bone) that are comprised of mineralized collagen fibrils and extrafibrillar matrix. The mechanical behavior of mineralized collagen fibrils has been investigated intensively using both experimental and computational approaches. Yet, the contribution of the extrafibrillar matrix to bone mechanical properties is poorly documented. In this study, we intended to address this issue using a novel cohesive finite element (FE) model, in conjunction with the experimental observations reported in the literature. In the FE model, the extrafibrillar matrix was considered as a nanocomposite of hydroxyapatite (HA) crystals bounded through a thin organic interface modeled as a cohesive interfacial zone. The parameters required by the cohesive FE model were defined based on the experimental data reported in the literature. This hybrid nanocomposite model was tested in two loading modes (i.e. tension and compression) and under two hydration conditions (i.e. wet and dry). The simulation results indicated that (1) the failure modes of the extrafibrillar matrix predicted using the cohesive FE model were closely coincided with those experimentally observed in tension and compression tests; (2) the pre-yield deformation (i.e. internal strain) of HA crystals with respect to the applied strain was consistent with that obtained from the synchrotron X-ray scattering measurements irrespective of the loading modes and hydration status; and (3) the mechanical behavior of the extrafibrillar matrix was dictated by the properties of the organic interface between the HA crystals. Taken together, we postulate that the extrafibrillar matrix plays a major role in the pre-yield deformation and the failure mode of bone, thus, giving rise to important insights in the ultrastructural origins of bone fragility., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Purpose: This double blind randomized clinical trial was conducted with the purpose of evaluating the effects of Nano-hydroxyapatite toothpaste as compared to 8% Arginine containing toothpaste in the management of Dentin hypersensitivity (DH)., Method and Materials: Patients (30 in each group) suffering from DH and eliciting a VAS score higher than 2 in air blast and tactile test were randomly allocated (block randomization) into either a group 1 (arginine toothpaste) or group 2 (nHA toothpaste). The primary outcome evaluated was the reduction of DH as measured by the electrical stimulus reading on the digital pulp tester. Current required for eliciting a VAS score of 2 was recorded before application of dentifrice. 1 cm of toothpaste was then expressed on the tooth surface for two minutes in each group and rinsed off. The electrical stimulus required to elicit a VAS score of 2 was recorded after 5 minutes, 1 week and 4 weeks., Results: The desensitizing paste containing arginine provided a statistically significant reduction in DH and so did the paste containing nHA. Mean increase in amperage value (reduction in DH) was higher for nHA based than the arginine containing dentifrice. This difference was not statistically significant showing that both toothpastes are equally effective., Conclusions: The findings of the present study encourage the use of Nano-hydroxyapatite and arginine containing dentifrice as an effective desensitizing agent providing relief from symptoms 5 minutes after application and after 1 and 4 weeks.

Dental enamel, an avascular, irreparable, outermost and protective layer of the human clinical crown has a potential to withstand the physico-chemical effects and forces. These properties are being regulated by a unique association among elements occurring in the crystallites setup of human dental enamel. Calcium and phosphate are the major components (hydroxyapatite) in addition to some trace elements which have a profound effect on enamel. The current review was planned to determine the aptitude of various trace elements to substitute and their influence on human dental enamel in terms of physical and chemical properties.