Hydrogel-Inducing Graphene-Oxide-Derived Core-Shell Fiber Composite for Antibacterial Wound Dressing.

The study reveals the polymer-crosslinker interactions and functionality of hydrophilic nanofibers for antibacterial wound coatings. Coaxial electrospinning leverages a drug encapsulation protocol for a core-shell fiber composite with a core derived from polyvinyl alcohol and polyethylene glycol with amorphous silica (PVA-PEG-SiO2), and a shell originating from polyvinyl alcohol and graphene oxide (PVA-GO). Crosslinking with GO and SiO2 initiates the hydrogel transition for the fiber composite upon contact with moisture, which aims to optimize the drug release. The effect of hydrogel-inducing additives on the drug kinetics is evaluated in the case of chlorhexidine digluconate (CHX) encapsulation in the core of core-shell fiber composite PVA-PEG-SiO2-1x-CHX@PVA-GO. The release rate is assessed with the zero, first-order, Higuchi, and Korsmeyer-Peppas kinetic models, where the inclusion of crosslinking silica provides a longer degradation and release rate. CHX medicated core-shell composite provides sustainable antibacterial activity against Staphylococcus aureus.

The Analysis of Micro-Scale Deformation and Fracture of Carbonized Elastomer-Based Composites by In Situ SEM.

Carbonized elastomer-based composites (CECs) possess a number of attractive features in terms of thermomechanical and electromechanical performance, durability in aggressive media and facile net-shape formability, but their relatively low ductility and strength limit their suitability for structural engineering applications. Prospective applications such as structural elements of micro-electro-mechanical systems MEMS can be envisaged since smaller principal dimensions reduce the susceptibility of components to residual stress accumulation during carbonization and to brittle fracture in general. We report the results of in situ in-SEM study of microdeformation and fracture behavior of CECs based on nitrile butadiene rubber (NBR) elastomeric matrices filled with carbon and silicon carbide. Nanostructured carbon composite materials were manufactured via compounding of elastomeric substance with carbon and SiC fillers using mixing rolling mill, vulcanization, and low-temperature carbonization. Double-edge notched tensile (DENT) specimens of vulcanized and carbonized elastomeric composites were subjected to in situ tensile testing in the chamber of the scanning electron microscope (SEM) Tescan Vega 3 using a Deben microtest 1 kN tensile stage. The series of acquired SEM images were analyzed by means of digital image correlation (DIC) using Ncorr open-source software to map the spatial distribution of strain. These maps were correlated with finite element modeling (FEM) simulations to refine the values of elastic moduli. Moreover, the elastic moduli were derived from unloading curve nanoindentation hardness measurements carried out using a NanoScan-4D tester and interpreted using the Oliver-Pharr method. Carbonization causes a significant increase of elastic moduli from 0.86 ± 0.07 GPa to 14.12 ± 1.20 GPa for the composite with graphite and carbon black fillers. Nanoindentation measurements yield somewhat lower values, namely, 0.25 ± 0.02 GPa and 9.83 ± 1.10 GPa before and after carbonization, respectively. The analysis of fractography images suggests that crack initiation, growth and propagation may occur both at the notch stress concentrator or relatively far from the notch. Possible causes of such response are discussed, namely, (1) residual stresses introduced by processing; (2) shape and size of fillers; and (3) the emanation and accumulation of gases in composites during carbonization.

Advanced Time-Stepping Interpretation of Fly-Scan Continuous Rotation Synchrotron Tomography of Dental Enamel Demineralization.

High-resolution spatial and temporal analysis and 3D visualization of time-dependent processes, such as human dental enamel acid demineralization, often present a challenging task. Overcoming this challenge often requires the development of special methods. Dental caries remains one of the most important oral diseases that involves the demineralization of hard dental tissues as a consequence of acid production by oral bacteria. Enamel has a hierarchically organized architecture that extends down to the nanostructural level and requires high resolution to study its evolution in detail. Enamel demineralization is a dynamic process that is best investigated with the help of in situ experiments. In previous studies, synchrotron tomography was applied to study the 3D enamel structure at certain time points (time-lapse tomography). Here, another distinct approach to time-evolving tomography studies is presented, whereby the sample image is reconstructed as it undergoes continuous rotation over a virtually unlimited angular range. The resulting (single) data set contains the data for multiple (potentially overlapping) intermediate tomograms that can be extracted and analyzed as desired using time-stepping selection of data subsets from the continuous fly-scan recording. One of the advantages of this approach is that it reduces the amount of time required to collect an equivalent number of single tomograms. Another advantage is that the nominal time step between successive reconstructions can be significantly reduced. We applied this approach to the study of acidic enamel demineralization and observed the progression of demineralization over time steps significantly smaller than the total acquisition time of a single tomogram, with a voxel size smaller than 0.5 µm. It is expected that the approach presented in this paper can be useful for high-resolution studies of other dynamic processes and for assessing small structural modifications in evolving hierarchical materials.

The DIAD Approach to Correlative Synchrotron X-ray Imaging and Diffraction Analysis of Human Enamel.

The Dual Imaging and Diffraction (DIAD) beamline at Diamond Light Source (Didcot, U.K.) implements a correlative approach to the dynamic study of materials based on concurrent analysis of identical sample locations using complementary X-ray modalities to reveal structural detail at various length scales. Namely, the underlying beamline principle and its practical implementation allow the collocation of chosen regions within the sample and their interrogation using real-space imaging (radiography and tomography) and reciprocal space scattering (diffraction). The switching between the two principal modes is made smooth and rapid by design, so that the data collected is interlaced to obtain near-simultaneous multimodal characterization. Different specific photon energies are used for each mode, and the interlacing of acquisition steps allows conducting static and dynamic experiments. Building on the demonstrated realization of this state-of-the-art approach requires further refining of the experimental practice, namely, the methods for gauge volume collocation under different modes of beam-sample interaction. To address this challenge, experiments were conducted at DIAD devoted to the study of human dental enamel, a hierarchical structure composed of hydroxyapatite mineral nanocrystals, as a static sample previously affected by dental caries (tooth decay) as well as under dynamic conditions simulating the process of acid demineralization. Collocation and correlation were achieved between WAXS (wide-angle X-ray scattering), 2D (radiographic), and 3D (tomographic) imaging. While X-ray imaging in 2D or 3D modes reveals real-space details of the sample microstructure, X-ray scattering data for each gauge volume provided statistical nanoscale and ultrastructural polycrystal reciprocal-space information such as phase and preferred orientation (texture). Careful registration of the gauge volume positions recorded during the scans allowed direct covisualization of the data from two modalities. Diffraction gauge volumes were identified and visualized within the tomographic data sets, revealing the underlying local information to support the interpretation of the diffraction patterns. The present implementation of the 4D microscopy paradigm allowed following the progression of demineralization and its correlation with time-dependent WAXS pattern evolution in an approach that is transferable to other material systems.

Hierarchical modelling of elastic behaviour of human enamel based on synchrotron diffraction characterisation.

Human enamel is a hierarchical mineralized tissue with a two-level composite structure. Few studies have focused on the structure-mechanical property relationship and its link to the multi-scale architecture of human enamel, whereby the response to mechanical loading is affected not only by the rod distribution at micro-scale, but also strongly influenced by the mineral crystallite shape, and spatial arrangement and orientation. In this study, two complementary synchrotron X-ray diffraction techniques, wide and small angle X-ray scattering (WAXS/SAXS) were used to obtain multi-scale quantitative information about the structure and deformation response of human enamel to in situ uniaxial compressive loading. The apparent modulus was determined linking the external load and the internal strain in hydroxyapatite (HAp) crystallites. An improved multi-scale Eshelby model is proposed taking into account the two-level hierarchical structure of enamel. This framework has been used to analyse the experimental data for the elastic lattice strain evolution within the HAp crystals. The achieved agreement between the model prediction and experiment along the loading direction validates the model and suggests that the new multi-scale approach reasonably captures the structure-property relationship for the human enamel. The ability of the model to predict multi-directional strain components is also evaluated by comparison with the measurements. The results are useful for understanding the intricate relationship between the hierarchical structure and the mechanical properties of enamel, and for making predictions of the effect of structural alterations that may occur due to the disease or treatment on the performance of dental tissues and their artificial replacements.

Synchrotron X-ray Studies of the Structural and Functional Hierarchies in Mineralised Human Dental Enamel: A State-of-the-Art Review.

Hard dental tissues possess a complex hierarchical structure that is particularly evident in enamel, the most mineralised substance in the human body. Its complex and interlinked organisation at the Ångstrom (crystal lattice), nano-, micro-, and macro-scales is the result of evolutionary optimisation for mechanical and functional performance: hardness and stiffness, fracture toughness, thermal, and chemical resistance. Understanding the physical-chemical-structural relationships at each scale requires the application of appropriately sensitive and resolving probes. Synchrotron X-ray techniques offer the possibility to progress significantly beyond the capabilities of conventional laboratory instruments, i.e., X-ray diffractometers, and electron and atomic force microscopes. The last few decades have witnessed the accumulation of results obtained from X-ray scattering (diffraction), spectroscopy (including polarisation analysis), and imaging (including ptychography and tomography). The current article presents a multi-disciplinary review of nearly 40 years of discoveries and advancements, primarily pertaining to the study of enamel and its demineralisation (caries), but also linked to the investigations of other mineralised tissues such as dentine, bone, etc. The modelling approaches informed by these observations are also overviewed. The strategic aim of the present review was to identify and evaluate prospective avenues for analysing dental tissues and developing treatments and prophylaxis for improved dental health.

Time-Lapse In Situ 3D Imaging Analysis of Human Enamel Demineralisation Using X-ray Synchrotron Tomography.

Caries is a chronic disease that causes the alteration of the structure of dental tissues by acid dissolution (in enamel, dentine and cementum) and proteolytic degradation (dentine and cementum) and generates an important cost of care. There is a need to visualise and characterise the acid dissolution process on enamel due to its hierarchical structure leading to complex structural modifications. The process starts at the enamel surface and progresses into depth, which necessitates the study of the internal enamel structure. Artificial demineralisation is usually employed to simulate the process experimentally. In the present study, the demineralisation of human enamel was studied using surface analysis carried out with atomic force microscopy as well as 3D internal analysis using synchrotron X-ray tomography during acid exposure with repeated scans to generate a time-lapse visualisation sequence. Two-dimensional analysis from projections and virtual slices and 3D analysis of the enamel mass provided details of tissue changes at the level of the rods and inter-rod substance. In addition to the visualisation of structural modifications, the rate of dissolution was determined, which demonstrated the feasibility and usefulness of these techniques. The temporal analysis of enamel demineralisation is not limited to dissolution and can be applied to other experimental conditions for the analysis of treated enamel or remineralisation.

Multi-resolution Correlative Ultrastructural and Chemical Analysis of Carious Enamel by Scanning Microscopy and Tomographic Imaging.

Caries, a major global disease associated with dental enamel demineralization, remains insufficiently understood to devise effective prevention or minimally invasive treatment. Understanding the ultrastructural changes in enamel is hampered by a lack of nanoscale characterization of the chemical spatial distributions within the dental tissue. This leads to the requirement to develop techniques based on various characterization methods. The purpose of the present study is to demonstrate the strength of analytic methods using a correlative technique on a single sample of human dental enamel as a specific case study to test the accuracy of techniques to compare regions in enamel. The science of the different techniques is integrated to genuinely study the enamel. The hierarchical structures within carious tissue were mapped using the combination of focused ion beam scanning electron microscopy with synchrotron X-ray tomography. The chemical changes were studied using scanning X-ray fluorescence (XRF) and X-ray wide-angle and small-angle scattering using a beam size below 80 nm for ångström and nanometer length scales. The analysis of XRF intensity gradients revealed subtle variations of Ca intensity in carious samples in comparison with those of normal mature enamel. In addition, the pathways for enamel rod demineralization were studied using X-ray ptychography. The results show the chemical and structural modification in carious enamel with differing locations. These results reinforce the need for multi-modal approaches to nanoscale analysis in complex hierarchically structured materials to interpret the changes of materials. The approach establishes a meticulous correlative characterization platform for the analysis of biomineralized tissues at the nanoscale, which adds confidence in the interpretation of the results and time-saving imaging techniques. The protocol demonstrated here using the dental tissue sample can be applied to other samples for statistical study and the investigation of nanoscale structural changes. The information gathered from the combination of methods could not be obtained with traditional individual techniques.

Acid-induced demineralisation of human enamel as a function of time and pH observed using X-ray and polarised light imaging.

Acid-induced enamel demineralisation affects many individuals either by exposure to acidic diets, acidic gas pollution (dental erosion) or to dental plaque acids (dental caries). This study aimed to develop in situ X-ray and light imaging methods to determine progression of enamel demineralisation and the dynamic relationship between acid pH and mineral density. Hourly digital microradiograph time-lapse sequences showed the depth of enamel demineralisation in 500 µm thick sections progressed with time from the surface towards the dentine following a power-law function, which was 21% faster than the lateral demineralisation progression after exposure for 85 h to lactic acid (10%, pH 2.2). The minimum greyscale remaining (mineral content) within the induced enamel lesion followed an exponential decay, while the accumulated total greyscale loss with time was linear, which showed a constant anisotropic mineral release within the enamel architecture. This 85 h demineralisation method studied by polarised light microscopy time-lapse sequences showed that once the demineralisation front reached the enamel Hunter-Schreger bands, there was preferential demineralisation along those bands. Mineral density loss was linear with increasing pH acidity between pH 5.2 and pH 4.0 (with 0.4 pH increments) when incubated over a 3-week period exposed to 0.5% lactic acid. At pH 4.0, there was complete mineral loss in the centre of the demineralised area after the 3-week period and the linear function intercepted the x-axis at ~ pH 5.5, near the critical pH for hydroxyapatite (HAp). These observations showed how intrinsic enamel structure and pH affected the progression of demineralisation. STATEMENT OF SIGNIFICANCE: Hydroxyapatite crystallites (HAp) in human enamel dissolve when exposed to an acidic environment but little is known about how the intrinsic structures in enamel and pH influence the demineralisation kinetics. We have developed a time-lapse in situ microradiography method to quantify microscopic anisotropic mineral loss dynamics in response to an acid-only caries model. Correlation with polarised light microscopy time-lapse sequences showed that larger structures in enamel also influence demineralisation progression as demineralisation occurred preferentially along the Hunter-Schreger bands (decussating prismatic enamel). The pH-controlled enamel mineral release in a linear manner quantifying the relationship between HAp orientation and acid solubility. These findings should direct the development of improved anti-demineralisation/ remineralisation treatments to retain/ restore the natural intrinsic enamel structure.

Investigations into the interface failure of yttria partially stabilised zirconia - porcelain dental prostheses through microscale residual stress and phase quantification.

OBJECTIVES: Yttria Partially Stabilised Zirconia (YPSZ) is a high strength ceramic which has become widely used in porcelain veneered dental copings due to its exceptional toughness. Within these components the residual stress and crystallographic phase of YPSZ close to the interface are highly influential in the primary failure mode; near interface porcelain chipping. In order to improve present understanding of this behaviour, characterisation of these parameters is needed at an improved spatial resolution. METHODS: In this study transmission micro-focus X-ray Diffraction, Raman spectroscopy, and focused ion beam milling residual stress analysis techniques have, for the first time, been used to quantify and cross-validate the microscale spatial variation of phase and residual stress of YPSZ in a prosthesis cross-section. RESULTS: The results of all techniques were found to be comparable and complementary. Monoclinic YPSZ was observed within the first 10µm of the YPSZ-porcelain interface with a maximum volume fraction of 60%. Tensile stresses were observed within the first 150 µm of the interface with a maximum value of ≈300 MPa at 50 µm from the interface. The remainder of the coping was in mild compression at ≈-30MPa, with shear stresses of a similar magnitude also being induced by the YPSZ phase transformation. SIGNIFICANCE: The analysis indicates that the interaction between phase transformation, residual stress and porcelain creep at YPSZ-porcelain interface results in a localised porcelain fracture toughness reduction. This explains the increased propensity of failure at this location, and can be used as a basis for improving prosthesis design.

Residual strain mapping through pair distribution function analysis of the porcelain veneer within a yttria partially stabilised zirconia dental prosthesis.

OBJECTIVE: Residually strained porcelain is influential in the early onset of failure in Yttria Partially Stabilised Zirconia (YPSZ) - porcelain dental prosthesis. In order to improve current understanding it is necessary to increase the spatial resolution of residual strain analysis in these veneers. METHODS: Few techniques exist which can resolve residual stress in amorphous materials at the microscale resolution required. For this reason, recent developments in Pair Distribution Function (PDF) analysis of X-ray diffraction data of dental porcelain have been exploited. This approach has facilitated high-resolution (70µm) quantification of residual strain in a YPSZ-porcelain dental prosthesis. In order to cross-validate this technique, the sequential ring-core focused ion beam and digital image correlation approach was implemented at a step size of 50µm. This semi-destructive technique exploits microscale strain relief to provide quantitative estimates of the near-surface residual strain. RESULTS: The two techniques were found to show highly comparable results. The residual strain within the veneer was found to be primarily tensile, with the highest magnitude stresses located at the YPSZ-porcelain interface where failure is known to originate. Oscillatory tensile and compressive stresses were also found in a direction parallel to the interface, likely to be induced by the multiple layering used during fabrication. SIGNIFICANCE: This study provides the insights required to improve prosthesis modelling, to develop new processing routes that minimise residual stress and ultimately to reduce prosthesis failure rates. The PDF approach also offers a powerful new technique for microscale strain quantification in amorphous materials.

In situ monitoring and analysis of enamel demineralisation using synchrotron X-ray scattering.

Dental caries is one of the most common chronic diseases that affect human teeth. It often initiates in enamel, undermining its mechanical function and structural integrity. Little is known about the enamel demineralisation process caused by dental caries in terms of the microstructural changes and crystallography of the inorganic mineral phase. To improve the understanding of the carious lesion formation process and to help identify efficient treatments, the evolution of the microstructure at the nano-scale in an artificially induced enamel erosion region was probed using advanced synchrotron small-angle and wide-angle X-ray scattering (SAXS and WAXS). This is the first in vitro and time-resolved investigation of enamel demineralisation using synchrotron X-ray techniques which allows in situ quantification of the microstructure evolution over time in a simulated carious lesion. The analysis revealed that alongside the reduction of mineral volume, a heterogeneous evolution of hydroxyapatite (HAp) crystallites (in terms of size, preferred orientation and degree of alignment) could be observed. It was also found that the rate and direction of dissolution depends on the crystallographic orientation. Based on these findings, a novel conceptual view of the process is put forward that describes the key structural parameters in establishing high fidelity ultrastructure-based numerical models for the simulation of the enamel demineralisation process. STATEMENT OF SIGNIFICANCE: Hydroxyapatite (HAp) crystallites in the enamel dissolve during dental caries although little is known about the structural-chemical relationships that control the dynamic demineralisation process. For the first time this work investigated the in situ evolution of nano-scale morphology and the spatial distribution of ultrastructural HAp crystallites of human enamel during demineralisation in simulated caries. Advanced synchrotron SAXS and WAXS techniques showed that the heterogeneous evolution of crystallites (size, preferred orientation and degree of alignment) could be attributed to crystallographic-orientation-dependent anisotropic dissolution. Hence we propose a novel conceptual schematic diagram to describe the demineralisation process. These findings have important implications for understanding the detailed mechanisms of enamel demineralisation and provide insight into potential enamel remineralisation that could restore structural integrity and function.

Structure-Function Correlative Microscopy of Peritubular and Intertubular Dentine.

Peritubular dentine (PTD) and intertubular dentine (ITD) were investigated by 3D correlative Focused Ion Beam (FIB)-Scanning Electron Microscopy (SEM)-Energy Dispersive Spectroscopy (EDS) tomography, tapping mode Atomic Force Microscopy (AFM) and scattering-type Scanning Near-Field Optical Microscopy (s-SNOM) mapping. The brighter appearance of PTD in 3D SEM-Backscattered-Electron (BSE) imaging mode and the corresponding higher grey value indicate a greater mineral concentration in PTD (~160) compared to ITD (~152). However, the 3D FIB-SEM-EDS reconstruction and high resolution, quantitative 2D map of the Ca/P ratio (~1.8) fail to distinguish between PTD and ITD. This has been further confirmed using nanoscale 2D AFM map, which clearly visualised biopolymers and hydroxyapatite (HAp) crystallites with larger mean crystallite size in ITD (32 ± 8 nm) than that in PTD (22 ± 3 nm). Correlative microscopy reveals that the principal difference between PTD and ITD arises primarily from the nanoscale packing density of the crystallites bonded together by thin biopolymer, with moderate contribution from the chemical composition difference. The structural difference results in the mechanical properties variation that is described by the parabolic stiffness-volume fraction correlation function introduced here. The obtained results benefit a microstructure-based mechano-chemical model to simulate the chemical etching process that can occur in human dental caries and some of its treatments.

Characterisation of nanovoiding in dental porcelain using small angle neutron scattering and transmission electron microscopy.

OBJECTIVES: Recent studies of the yttria partially stabilised zirconia-porcelain interface have revealed the presence of near-interface porcelain nanovoiding which reduces toughness and leads to component failure. One potential explanation for these nanoscale features is thermal creep which is induced by the combination of the residual stresses at the interface and sintering temperatures applied during manufacture. The present study provides improved understanding of this important phenomenon. METHODS: Transmission electron microscopy and small angle neutron scattering were applied to a sample which was crept at 750°C and 100MPa (sample C), a second which was exposed to an identical heat treatment schedule in the absence of applied stress (sample H), and a reference sample in the as-machined state (sample A). RESULTS: The complementary insights provided by the two techniques were in good agreement and log-normal void size distributions were found in all samples. The void number density was found to be 1.61µm-2, 25.4µm-2 and 98.6µm-2 in samples A, H and C respectively. The average void diameter in sample A (27.1nm) was found to be more than twice as large as in samples H (10.2nm) and C (11.6nm). The crept data showed the highest skewness parameter (2.35), indicating stress-induced growth of larger voids and void coalescence that has not been previously observed. SIGNIFICANCE: The improved insight presented in this study can be integrated into existing models of dental prostheses in order to optimise manufacturing routes and thereby reduce the significant detrimental impact of this nanostructural phenomenon.

Understanding nature's residual strain engineering at the human dentine-enamel junction interface.

Human dental tissue is a hydrated biological mineral composite. In terms of volume and mass, a human tooth mainly consists of dentine and enamel. Human dental tissues have a hierarchical structure and versatile mechanical properties. The dentine enamel junction (DEJ) is an important biological interface that provides a durable bond between enamel and dentine that is a life-long success story: while intact and free from disease, this interface does not fail despite the harsh thermo-mechanical loading in the oral cavity. The underlying reasons for such remarkable strength and durability are still not fully clear from the structural and mechanical perspectives. One possibility is that, in an example of residual stress engineering, evolution has led to the formation of a layer of inelastic strain adjacent to the DEJ during odontogenesis (tooth formation). However, due to significant experimental and interpretational challenges, no meaningful quantification of residual stress in the vicinity of the DEJ at the appropriate spatial resolution has been reported to date. In this study, we applied a recently developed flexible and versatile method for measuring the residual elastic strain at (sub)micron-scale utilising focused ion beam (FIB) milling with digital image correlation (DIC). We report the results that span the transition from human dentine to enamel, and incorporate the material lying at and in the vicinity of the DEJ. The capability of observing the association between internal architecture and the residual elastic strain state at the micrometre scale is useful for understanding the remarkable performance of the DEJ and may help the creation of improved biomimetic materials for clinical and engineering applications. STATEMENT OF SIGNIFICANCE: We studied the micron-scale residual stresses that exist within human teeth, between enamel (outer tooth shell, hardest substance in the human body) and dentine (soft bone-like vascularised tooth core). The dentine-enamel junction (DEJ) is an extremely interesting example of nature's design in terms of hierarchical structuring and residual stress management. Key developments reported are systematic focused ion beam (FIB) milling and digital image correlation (DIC) micrometre scale residual strain evaluation, and the determination of principal strain direction near DEJ, correlated with internal architecture responsible for remarkable strength. This work helps understanding DEJ performance and improving biomimetic materials design for clinical and engineering applications.

X-ray scattering evaluation of ultrastructural changes in human dental tissues with thermal treatment.

Micro- and ultrastructural analysis of burned skeletal remains is crucial for obtaining a reliable estimation of cremation temperature. Earlier studies mainly focused on heat-induced changes in bone tissue, while this study extends this research to human dental tissues using a novel quantitative analytical approach. Twelve tooth sections were burned at 400-900°C (30-min exposure, increments of 100°C). Subsequent combined small- and wide-angle X-ray scattering (SAXS/WAXS) experiments were performed at the Diamond Light Source synchrotron facility, where 28 scattering patterns were collected within each tooth section. In comparison with the control sample, an increase in mean crystal thickness was found in burned dentine (2.8-fold) and enamel (1.4-fold), however at a smaller rate than reported earlier for bone tissue (5-10.7-fold). The results provide a structural reference for traditional X-ray scattering methods and emphasize the need to investigate bone and dental tissues separately to obtain a reliable estimation of cremation temperature.

Structure-mechanical function relations at nano-scale in heat-affected human dental tissue.

The knowledge of the mechanical properties of dental materials related to their hierarchical structure is essential for understanding and predicting the effect of microstructural alterations on the performance of dental tissues in the context of forensic and archaeological investigation as well as laser irradiation treatment of caries. So far, few studies have focused on the nano-scale structure-mechanical function relations of human teeth altered by chemical or thermal treatment. The response of dental tissues to thermal treatment is thought to be strongly affected by the mineral crystallite size, their spatial arrangement and preferred orientation. In this study, synchrotron-based small and wide angle X-ray scattering (SAXS/WAXS) techniques were used to investigate the micro-structural alterations (mean crystalline thickness, crystal perfection and degree of alignment) of heat-affected dentine and enamel in human dental teeth. Additionally, nanoindentation mapping was applied to detect the spatial and temperature-dependent nano-mechanical properties variation. The SAXS/WAXS results revealed that the mean crystalline thickness distribution in dentine was more uniform compared with that in enamel. Although in general the mean crystalline thickness increased both in dentine and enamel as the temperature increased, the local structural variations gradually reduced. Meanwhile, the hardness and reduced modulus in enamel decreased as the temperature increased, while for dentine, the tendency reversed at high temperature. The analysis of the correlation between the ultrastructure and mechanical properties coupled with the effect of temperature demonstrates the effect of mean thickness and orientation on the local variation of mechanical property. This structural-mechanical property alteration is likely to be due to changes of HAp crystallites, thus dentine and enamel exhibit different responses at different temperatures. Our results enable an improved understanding of the mechanical properties correlation in hierarchical biological materials, and human dental tissue in particular.

In situ X-ray scattering evaluation of heat-induced ultrastructural changes in dental tissues and synthetic hydroxyapatite.

Human dental tissues consist of inorganic constituents (mainly crystallites of hydroxyapatite, HAp) and organic matrix. In addition, synthetic HAp powders are frequently used in medical and chemical applications. Insights into the ultrastructural alterations of skeletal hard tissues exposed to thermal treatment are crucial for the estimation of temperature of exposure in forensic and archaeological studies. However, at present, only limited data exist on the heat-induced structural alterations of human dental tissues. In this paper, advanced non-destructive small- and wide angle X-ray scattering (SAXS/WAXS) synchrotron techniques were used to investigate the in situ ultrastructural alterations in thermally treated human dental tissues and synthetic HAp powders. The crystallographic properties were probed by WAXS, whereas HAp grain size distribution changes were evaluated by SAXS. The results demonstrate the important role of the organic matrix that binds together the HAp crystallites in responding to heat exposure. This is highlighted by the difference in the thermal behaviour between human dental tissues and synthetic HAp powders. The X-ray analysis results are supported by thermogravimetric analysis. The results concerning the HAp crystalline architecture in natural and synthetic HAp powders provide a reliable basis for deducing the heating history for dental tissues in the forensic and archaeological context, and the foundation for further development and optimization of biomimetic material design.

Multiscale modelling and diffraction-based characterization of elastic behaviour of human dentine.

Human dentine is a hierarchical mineralized tissue with a two-level composite structure, with tubules being the prominent structural feature at a microlevel, and collagen fibres decorated with hydroxyapatite (HAp) crystallite platelets dominating the nanoscale. Few studies have focused on this two-level structure of human dentine, where the response to mechanical loading is thought to be affected not only by the tubule volume fraction at the microscale, but also by the shape and orientation distribution of mineral crystallites, and their nanoscale spatial arrangement and alignment. In this paper, in situ elastic strain evolution within HAp in dentine subjected to uniaxial compressive loading along both longitudinal and transverse directions was characterized simultaneously by two synchrotron X-ray scattering techniques: small- and wide-angle X-ray scattering (SAXS and WAXS, respectively). WAXS allows the evaluation of the apparent modulus linking the external load to the internal HAp crystallite strain, while the nanoscale HAp distribution and arrangement can be quantified by SAXS. We proposed an improved multiscale Eshelby inclusion model that takes into account the two-level hierarchical structure, and validated it with a multidirectional experimental strain evaluation. The agreement between the simulation and measurement indicates that the multiscale hierarchical model developed here accurately reflects the structural arrangement and mechanical response of human dentine. This study benefits the comprehensive understanding of the mechanical behaviour of hierarchical biomaterials. The knowledge of the mechanical properties related to the hierarchical structure is essential for the understanding and predicting the effects of structural alterations that may occur due to disease or treatment on the performance of dental tissues and their artificial replacements.