Intraoral low-temperature degradation of monolithic zirconia dental prostheses: 5-year results of a prospective clinical study with ex vivo monitoring.

OBJECTIVES: To investigate the 5-year intraoral evolution and kinetics of low-temperature degradation (LTD) of second-generation monolithic prostheses made of 3% molar yttrium-doped tetragonal zirconia polycrystal (3Y-TZP) and the influence of masticatory mechanical stresses and glaze layer on this evolution. METHODS: A total of 101 posterior tooth elements were included in this prospective clinical study, which comprised ex vivo LTD monitoring (at baseline, 6 months, 1 year, 2 years, 3 years, and 5 years) using Raman spectroscopy (n = 2640 monoclinic phase measurement points per evaluation time) and scanning electron microscopy (SEM). Four types of areas (1-2 mm2 surface, six on molars, and four on premolars) were analysed on each element surface: occlusal, axial, glazed, or unglazed. Raman mapping, high-resolution SEM, and focused ion beam-SEM were performed on selected samples. RESULTS: The dental prostheses developed a tetragonal-to-monoclinic transformation at the extreme surface of the material after six months in a buccal environment, and this process increased significantly over time. Over the five years of monitoring, the transformation developed nonuniformly with the presence of localised clusters of monoclinic grains. Tribological stresses generate grain pull-out from these clusters, which may raise questions regarding the release of 3Y-TZP nanoparticles into the body. The prosthesis fracture rate was 4.5% after 5 years. SIGNIFICANCE: LTD developed in vivo on the surfaces of 3Y-TZP dental prostheses and progressed slowly but significantly over time, up to 5 years investigation. However, the effects of aging on the failure rate recorded and of zirconia nanoparticles released into the body require further investigation.

Intraoral low-temperature degradation of monolithic zirconia dental prostheses: Results of a prospective clinical study with ex vivo monitoring.

OBJECTIVE: To investigate the intraoral development and kinetics of low-temperature degradation (LTD) in second-generation 3 mol.% yttria-doped tetragonal zirconia polycrystal (3Y-TZP) monolithic prostheses, as well as the influence of masticatory mechanical stress and glaze layer on it. METHODS: A total of 101 posterior tooth elements were included in a prospective clinical study, which included ex vivo LTD monitoring (at baseline, 6 months, 1 year, and 2 years) using Raman spectroscopy (n = 2640 monoclinic phase measurement points per evaluation time) and SEM. Four types of areas (1-2 mm2 surface, 6 on molars, and 4 on premolars) were analyzed on each element surface: occlusal, axial, glazed, or unglazed. Raman depth mapping and high-resolution SEM were performed on the selected samples. RESULTS: LTD developed in 3Y-TZP monolithic restorations 6 months after intraoral placement and progressed with time. After two years, the tetragonal-to-monoclinic transformation was non-uniform, with the presence of localized clusters of transformed grains. In axial areas, the grain aspect was typical of the classical nucleation-growth process reported for LTD, which progresses from the surface to a depth of several tens of microns. However, in occlusal areas, tribological stress generated surface crushing and grain pull-out from the clusters, which induced an underestimation of the aging process when the evaluation was limited to monoclinic phase quantification. Glazing cannot be considered a protection against LTD. SIGNIFICANCE: If LTD occurs in dental prostheses in the same way as in orthopedic prostheses, its clinical impact is unknown and needs to be further studied.

Intrinsic properties of osteomalacia bone evaluated by nanoindentation and FTIRM analysis.

Osteomalacia is a pathological bone condition consisting in a deficient primary mineralization of the matrix, leading to an accumulation of osteoid tissue and reduced bone mechanical strength. The amounts, properties and organization of bone constituents at tissue level, are known to influence its mechanical properties. It is then important to investigate the relationship between mechanical behavior and tissue composition at this scale in order to provide a better understanding of bone fragility mechanisms associates with this pathology. Our purpose was to analyze the links between ultra-structural properties and the mechanical behavior of this pathological bone tissue (osteomalacia) at tissue level (mineral and osteoid separately, or global). Four bone biopsies were taken from patients with osteomalacia, and subsequently embedded, sectioned, and polished. Then nanoindentation tests were performed to determine local elastic modulus E, contact hardness Hc and true hardness H for both mineralized and organic bone phases and for the global bone. The creep of the bone was also studied using a special indentation procedure in order to assess visco-elasto-plastic (creep) bone behavior. This allowed a detailed study of the rheological models adapted to the bone and to calculate the parameters associated to a Burgers model. Ultra-structural parameters were measured by Fourier Transform InfraRed Microspectroscopy (FTIRM) on the same position as the indents. The use of rheological models confirmed a significant contribution from the organic phase on the viscous character of bone tissue. The elastic E and the elasto-plastic Hc deformation were correlated to both collagen maturity and Mineral/Matrix. The pure plastic deformation H was only correlated to the mineral phase. Our data show that mineral phase greatly affects mechanical variables (moduli and viscosities) and that organic phase (as illustrated in osteoid tissue) may play an important role in the creep behavior of bone. In conclusion, this study brings mechanical and physicochemical values for osteoid and mineral phases.

3D microstructural study of selachimorph enameloid evolution.

Enameloid, the hyper-mineralized tissue covering shark teeth is a complex structure resulting from both ameloblast and odontoblast activity. The way these two types of cells interact to set up this tissue is not fully understood and results in the formation of subunits in the enameloid: the Single Crystallite Enameloid (SCE) and the Bundled Crystallite Enameloid (BCE). Using the Focused Ion Beam Nanotomography (FIB-nt), 3D images were produced to assess the relationship between the SCE and BCE of one fossil and one recent neoselachian shark teeth. 3D analysis of crystallite bundles reveals a strong connection between the crystallites forming the SCE and those forming the bundles of the Radial Bundle Enameloid (RBE), a component of the BCE, although it has been suggested that SCE and BCE have a different origin: epithelial for the SCE and mesenchymal for the BCE. Another significant result of the use of FIB-nt is the visualization of frequent branching among the radial bundles forming the RBE, including horizontal link between adjacent bundles. FIB-nt demonstrates therefore a strong potential to decipher the complex evolution of hyper-mineralised tissue in shark teeth, and, therefore, to better understand the evolution of tooth structure among basal Gnathostomes.

Towards large scale orientation mapping using the eCHORD method.

eCHORD is an alternative method for orientation mapping in Scanning Electron Microscopy that involves channeling contrast observed in BSE images. The sample tilt being small (10°), eCHORD could be a promising method for large scale maps as it limits the image deformation sometimes observed with EBSD maps. However, when the magnification is low, the scan deflection of the beam becomes important, which may modify the channeling conditions of analyzed grains, leading to orientation errors. A correction method for the resulting orientations is proposed and a resulting map on a Si single crystal is presented with an experimental misorientation of 0.12° across a field of view of 2.3 × 1.7 mm2.

Angular resolution expected from iCHORD orientation maps through a revisited ion channeling model.

Crystalline orientation maps are obtained in a Focused Ion Beam (FIB) microscope using the ion CHanneling ORientation Determination (iCHORD) method, which relies on the channeling phenomenon observed in ion-induced secondary electron images. The current paper focuses on the angular resolution that can be expected from such orientation maps, obtained using a revisited ion channeling model. A specific procedure was developed to evaluate the angular resolution, based on the distribution of orientation errors when evaluating controlled sample disorientation. The main advantage is that no external reference is required. An angular resolution of 1° is obtained on a nickel based sample using standard acquisition conditions. This value fulfills most of the needs in terms of microstructural characterization usually carried out by Electron Back Scattered Diffraction.

Electron CHanneling ORientation Determination (eCHORD): An original approach to crystalline orientation mapping.

We present a proof-of-concept attesting the feasability to obtain orientation maps of polycrystalline materials within a conventional Scanning Electron Microscope (SEM) using a standard goniometer and Back Scattered Electron (BSE) detector. The described method is based on the analysis of the contrast variation of grains due to the channeling of incident electrons on a rotating sample. On each pixel of the map, experimental intensity profiles as a function of the rotation angle are obtained and compared with simulated ones to retrieve the orientation. From first results on aluminum polycrystals, the angular resolution is estimated to be better than one degree.

Sub-surface assessment of hydrothermal ageing in zirconia-containing femoral heads for hip joint applications.

Zirconia-based materials have been used in orthopaedics since the 1980s, with large success, mainly thanks to transformation toughening. On the other hand, their main drawback is their potential sensitivity to hydrothermal ageing, i.e. tetragonal to monoclinic phase transformation on their surface in the presence of water. Hydrothermal ageing may result in roughness increase and microcracking of the surface. In this article the hydrothermal ageing behaviour of three medical-grade zirconia-based materials is assessed at high temperature and extrapolated to room or body temperature. The degradation is also characterized by FIB/SEM nano-tomography to better assess sub-surface evolutions. In both zirconia and alumina-toughened zirconia (ATZ), ageing results in the presence of a homogenous transformed layer of constant thickness whose growth rate is about 8 times slower in ATZ than in zirconia. Microcracking occurs in the entire transformed layer in zirconia, but was much less relevant in ATZ. Zirconia-toughened alumina (ZTA) is much less prone to ageing. In ZTA ageing results in a thin transformed layer in which the monoclinic fraction decreases with depth. No microcracking was observed in ZTA. STATEMENT OF SIGNIFICANCE: This article details the microstructural evolution of the surface of three zirconia-based ceramics when exposed to water (hydrothermal ageing), and establishes a time-temperature equivalences of these evolutions. It shows that different zirconia-alumina composites do not degrade the same way: zirconia and alumina-toughened zirconia present a homogeneous degraded zone of constant thickness, whereas zirconia-toughened-alumina presents a gradient of transformation. These new findings will help understanding better the hydrothermal degradation of zirconia based materials, and in particular will facilitate a better prediction of the durability of zirconia-based devices such as orthopaedic implants and dental devices (implants, crowns, abutments…).

Nerve Growth Factor Expression and Its Receptors TrkA and p75NTR in Peri-Implantitis Lesions.

BACKGROUND: Nerve growth factor (NGF) can, through its receptors TrkA and p75NTR, convey signals for cell survival or cell differentiation. These proteins are also involved in inflammation and in bone resorption. The aim of this study is to evaluate, for the first time, the expression of NGF and its receptors TrkA and p75NTR in peri-implantitis lesions. MATERIALS AND METHODS: Fifteen biopsy specimens from patients with chronic peri-implantitis and 4 of healthy oral mucosa were immunostained with antibodies against NGF, TrkA, and p75NTR. The staining intensity and percentage of stained cells were semi-quantitatively evaluated and results were compared between the 2 groups. RESULTS: In the peri-implant pocket epithelium and gingival epithelium, NGF and TrkA expressions were similar to the healthy oral mucosa, however, a decreased expression of p75NTR was observed. In all cases, more than 75% of the inflammatory cells stained positively for NGF and TrkA, and p75NTR was negatively expressed. CONCLUSION: The intense expression of NGF and TrkA in the inflammatory cell infiltrate associated with decreased expression of p75NTR in both gingival and pocket epithelium suggests that these proteins may have a role in peri-implantitis lesions.

M23C6 carbides and Cr2N nitrides in aged duplex stainless steel: A SEM, TEM and FIB tomography investigation.

The precipitation evolution during ageing of a 2101 lean duplex stainless steel was investigated, revealing that the precipitate type and morphology depends on the nature of the grain boundary. Triangular M23C6 carbides precipitate only at γ/δ interfaces and rod-like Cr2N nitrides precipitate at both γ/δ and δ/δ interfaces. After 15min of ageing, the M23C6 size no longer evolves, whereas that of the Cr2N continues to evolve. For Cr2N, the morphology is maintained at γ/δ interfaces, whereas percolation occurs to form a continuous layer at δ/δ interfaces. By combining 2D and 3D characterisation at the nanoscale using transmission electron microscopy (TEM) and focused ion beam (FIB) tomography, a complete description of the precipitation evolution was obtained, including the composition, crystallographic structure, orientation relationship with the matrix phases, location, morphology, size and volume fraction.

Crystal orientation mapping via ion channeling: An alternative to EBSD.

A new method, which we name ion CHanneling ORientation Determination (iCHORD), is proposed to obtain orientation maps on polycrystals via ion channeling. The iChord method exploits the dependence between grain orientation and ion beam induced secondary electron image contrast. At each position of the region of interest, intensity profiles are obtained from a series of images acquired with different orientations with respect to the ion beam. The profiles are then compared to a database of theoretical profiles of known orientation. The Euler triplet associated to the most similar theoretical profile gives the orientation at that position. The proof-of-concept is obtained on a titanium nitride sample. The potentialities of iCHORD as an alternative to EBSD are then discussed.

Effect of different surface treatments on the hydrothermal degradation of a 3Y-TZP ceramic for dental implants.

OBJECTIVES: Implant surface modifications are intended to enhance bone integration. The objective of this study was to assess the effect of different surface treatments on the resistance to hydrothermal degradation, hardness and elastic modulus of a 3Y-TZP ceramic used for dental implants. METHODS: Samples grouped according to their surface morphologies (AS, as-sintered; C, coated; P, dry-polished; R, roughened; PA, polished and annealed; RA, roughened and annealed) were subjected to accelerated hydrothermal degradation (LTD) by exposure to water steam (134°C, 2bars) for 100h. The t-m phase transformation was quantified by grazing incidence X-ray diffraction (GIXDR) and by combined focused ion beam and scanning electron microscopy (FIB-SEM). Elastic modulus and hardness before- and after prolonged aging (100h) were assessed by nanoindentation. RESULTS: AS and C specimens presented a better resistance to hydrothermal degradation than P and R samples. After prolonged aging, the depth of the monoclinic transformed layer ranged from 11µm to 14µm. Hydrothermal degradation led to a significant decrease of elastic modulus and hardness. SIGNIFICANCE: Surface treatments affected the resistance to hydrothermal degradation of the 3Y-TZP ceramic. Dry mechanical surface modifications should be avoided since a high t-m transformation rate associated to the initial monoclinic content was observed. Annealing was useful to reverse the initial t-m transformation, but did not improve the resistance to hydrothermal degradation.

A global investigation into in situ nanoindentation experiments on zirconia: from the sample geometry optimization to the stress nanolocalization using convergent beam electron diffraction.

Nanoindentation experiments inside a transmission electron microscope are of much interest to characterize specific phenomena occuring in materials, like for instance dislocation movements or phase transformations. The key points of these experiments are (i) the sample preparation and the optimization of its geometry to obtain reliable results and (ii) the choice of the transmission electron microscope observation mode, which will condition the type of information which can be deduced from the experiment. In this paper, we will focus on these two key points in the case of nanoindentation of zirconia, which is a ceramic material well known to be sensitive to stress because it can undergo a phase transformation. In this case, the information sought is the stress localization at the nanometre scale and in real time. As far as the sample preparation is concerned, one major drawback of nanoindentation inside a transmission electron microscope is indeed a possible bending of the sample occurring during compression, which is detrimental to the experiment interpretation (the stress is not uniaxial anymore). In this paper, several sample preparation techniques have been used and compared to optimize the geometry of the sample to avoid bending. The results obtained on sample preparation can be useful for the preparation of ceramics samples but can also give interesting clues and experimental approaches to optimize the preparation of other kinds of materials. The second part of this paper is devoted to the second key point, which is the determination of the stress localization associated to the deformation phenomena observed by nanoindentation experiments. In this paper, the use of convergent beam electron diffraction has been investigated and this technique could have been successfully coupled to nanoindentation experiments. Coupled nanoindentation experiments and convergent beam electron diffraction analyses have finally been applied to characterize the phase transformation of zirconia.

Respective roles of organic and mineral components of human cortical bone matrix in micromechanical behavior: an instrumented indentation study.

Bone is a multiscale composite material made of both a type I collagen matrix and a poorly crystalline apatite mineral phase. Due to remodeling activity, cortical bone is made of Bone Structural Units (BSUs) called osteons. Since osteon represents a fundamental level of structural hierarchy, it is important to investigate the relationship between mechanical behavior and tissue composition at this scale for a better understanding of the mechanisms of bone fragility. The aim of this study is to analyze the links between ultrastructural properties and the mechanical behavior of bone tissue at the scale of osteon. Iliac bone biopsies were taken from untreated postmenopausal osteoporotic women, embedded, sectioned and microradiographed to assess the degree of mineralization of bone (DMB). On each section, BSUs of known DMB were indented with relatively high load (~500 mN) to determine local elastic modulus (E), contact hardness (H(c)) and true hardness (H) of several bone lamellae. Crystallinity and collagen maturity were measured by Fourier Transform InfraRed Microspectroscopy (FTIRM) on the same BSUs. Inter-relationships between mechanical properties and ultrastructural components were analyzed using multiple regression analysis. This study showed that elastic deformation was only explained by DMB whereas plastic deformation was more correlated with collagen maturity. Contact hardness, reflecting both elastic and plastic behaviors, was correlated with both DMB and collagen maturity. No relationship was found between crystallinity and mechanical properties at the osteon level.