Photometric calibration of an in situ broadband optical thickness monitoring of thin films in a large vacuum chamber.

To improve the in situ monitoring of thin films at the Laboratoire des Matériaux Avancés, a broadband optical monitoring of the coated thin films was developed and installed in the biggest ion-beam sputtering machine in the world. Due to the configuration of the coating machine and the chamber strain under vacuum, a standard calibration procedure is impossible and a double-beam optical system is not suitable. A novel theoretical and practical solution to calibrate the measurements was found and is described in this paper. Some relevant results achieved thanks to this technique are discussed as well.

Extraction and analysis of body-induced partials of guitar tones.

Guitar plucked sounds arise from a rapid input of energy applied to the string coupled to the instrument body at the bridge. For the radiated pressure, this results in quasi-harmonic contributions, reflecting the string modes coupled to the body, as well as some transient and quickly decaying components reflecting the excitation of the body modes of the instrument. In order to evaluate the relevance of this transient body sound, a high resolution analysis-synthesis method is described for the extraction of the body-mode contribution from the radiated pressure measured in the near field of the guitar top plate. This analysis scheme is first tested on synthesized signals. Some body-sound emergence indicators are then proposed and computed over a pool of instruments. The influence of the conditions of excitation on the body-sound emergence is investigated, and the instruments categorized according to these objective descriptors. Results show a larger range of body-sound emergence with variations of the plucking position in hand-made guitars compared to industrial instruments. This suggests that these particular hand-made instruments are more sensitive to variations in the control from the player and hence allow a wider range of timbres with respect to the transient coloration of the body modes.

Acoustic signature of violins based on bridge transfer mobility measurements.

This paper is an attempt to solve two problems related to musical acoustics. The first one consists in defining a signature of an instrument, namely, summarizing its vibroacoustical behavior. The second one deals with the existing relationship between the musical sound and the vibroacoustic properties of the instrument body. The violin is the application of this paper. A proposed solution for the first problem consists in an estimation of the bridge transfer mobility and the mean-value of the lateral bridge transfer mobility. The second problem is studied via the comparison between the amplitudes of harmonics, extracted from a glissando audio signal, and the lateral bridge transfer mobility: Both curves exhibit similar features. This is the main result of the paper. This is evidenced by studying the effect of a violin mute on both the lateral bridge transfer mobility and the produced sound. Finally, this is evidenced by successfully identifying which violin is played in an audio recording, using the computation of the Pearson distance between the distribution of the amplitude of harmonics and a database of measured mobilities.

Osseointegration of polyethylene implants coated with titanium and biomimetically or electrochemically deposited hydroxyapatite in a rabbit model.

PURPOSE: The aim of this study was to evaluate the osseointegration of a new coating directly deposited on PE at room temperature. METHODS: Thirty-six (36) male New Zealand rabbits were randomly assigned to receive one out of three types of implants: two tested implants, i.e. PE implant coated with TiPVD and biomimetic HA (biomimetic), PE implant coated with TiPVD and electrolytic HA (electrolytic), and positive control made of massive microrough titanium coated with plasma sprayed HA (TiHAPS). Osseointegration was evaluated by histomorphometry (bone tissue in contact [BIC]), mineralized bone area [MBA]) and mechanical testing (push-out test, interfacial shear strength [ISS]) at six and 12 weeks in the distal femurs. RESULTS: For BIC there were no differences between the groups at six (p = 0.98) and 12 weeks (p = 0.13). For MBA, no statistically significant difference was measured between groups at six (p = 0.52) and 12 weeks (p = 0.57). At six weeks, interfacial shear strength (ISS) was significantly higher (p = 0.01) for TiHAPs implants compared to biomimetic and electrolytic implants. This difference was not significant at 12 weeks (p = 0.92). CONCLUSION: The osseointegration of biomimetic and electrolytic implants was equivalent to a positive control at 12 weeks.

A parametric model and estimation techniques for the inharmonicity and tuning of the piano.

Inharmonicity of piano tones is an essential property of their timbre that strongly influences the tuning, leading to the so-called octave stretching. It is proposed in this paper to jointly model the inharmonicity and tuning of pianos on the whole compass. While using a small number of parameters, these models are able to reflect both the specificities of instrument design and tuner's practice. An estimation algorithm is derived that can run either on a set of isolated note recordings, but also on chord recordings, assuming that the played notes are known. It is applied to extract parameters highlighting some tuner's choices on different piano types and to propose tuning curves for out-of-tune pianos or piano synthesizers.

Perfusion of MC3T3E1 Preosteoblast Spheroids within Polysaccharide-Based Hydrogel Scaffolds: An Experimental and Numerical Study at the Bioreactor Scale.

The traditional 3D culture systems in vitro lack the biological and mechanical spatiotemporal stimuli characteristic to native tissue development. In our study, we combined porous polysaccharide-based hydrogel scaffolds with a bioreactor-type perfusion device that generates favorable mechanical stresses while enhancing nutrient transfers. MC3T3E1 mouse osteoblasts were seeded in the scaffolds and cultivated for 3 weeks under dynamic conditions at a perfusion rate of 10 mL min-1. The spatial distribution of the cells labeled with superparamagnetic iron oxide nanoparticles was visualized by MRI. Confocal microscopy was used to assess cell numbers, their distribution inside the scaffolds, cell viability, and proliferation. The oxygen diffusion coefficient in the hydrogel was measured experimentally. Numerical simulations of the flow and oxygen transport within the bioreactor were performed using a lattice Boltzmann method with a two-relaxation time scheme. Last, the influence of cell density and spheroid size on cell oxygenation was investigated. The cells spontaneously organized into spheroids with a diameter of 30-100 µm. Cell viability remained unchanged under dynamic conditions but decreased under static culture. The cell proliferation (Ki67 expression) in spheroids was not observed. The flow simulation showed that the local fluid velocity reached 27 mm s-1 at the height where the cross-sectional area of the flow was the smallest. The shear stress exerted by the fluid on the scaffolds may locally rise to 100 mPa, compared with the average value of 25 mPa. The oxygen diffusion coefficient in the hydrogel was 1.6×10-9 m2 s-1. The simulation of oxygen transport and consumption confirmed that the cells in spheroids did not suffer from hypoxia when the bioreactor was perfused at 10 mL min-1, and suggested the existence of optimal spheroid size and spacing for appropriate oxygenation. Collectively, these findings enabled us to define the optimal conditions inside the bioreactor for an efficient in vitro cell organization and survival in spheroids, which are paramount to future applications with organoids.

Macro parameters describing the mechanical behavior of classical guitars.

Since the 1960s and 1970s, researchers have proposed simplified models using only a few parameters to describe the vibro-acoustical behavior of string instruments in the low-frequency range. This paper presents a method for deriving and estimating a few important parameters or features describing the mechanical behavior of classical guitars over a broader frequency range. These features are selected under the constraint that the measurements may readily be made in the workshop of an instrument maker. The computations of these features use estimates of the modal parameters over a large frequency range, made with the high-resolution subspace ESPRIT algorithm (Estimation of Signal Parameters via Rotational Invariant Techniques) and the signal enumeration technique ESTER (ESTimation of ERror). The methods are applied to experiments on real metal and wood plates and numerical simulations of them. The results on guitars show a nearly constant mode density in the mid- and high-frequency ranges, as it is found for a flat panel. Four features are chosen as characteristic parameters of this equivalent plate: Mass, rigidity, characteristic admittance, and the mobility deviation. Application to a set of 12 guitars indicates that these features are good candidates to discriminate different classes of classical guitars.

Interplay between crosslinking and ice nucleation controls the porous structure of freeze-dried hydrogel scaffolds.

Freeze-drying is a process of choice to texture hydrogel scaffolds with pores formed by an ice-templating mechanism. Using state-of-the-art microscopies (cryo-EBSD, µCT, CLSM), this work evidences and quantifies the effect of crosslinking and ice nucleation temperature on the porous structure of thin hydrogel scaffolds freeze-dried at a low cooling rate. We focused on a polysaccharide-based hydrogel and developed specific protocols to monitor or trigger ice nucleation for this study. At a fixed number of intermolecular crosslinks per primary molecule (p = 5), the mean pore size in the dry state decreases linearly from 240 to 170 µm, when ice nucleation temperature decreases from -6 °C to -18 °C. When ice nucleation temperature is fixed at -10 °C, the mean pore size decreases from 250 to 150 µm, as the crosslinking degree increases from p = 3 to p = 7. Scaffold infiltration ability was quantified with synthetic microspheres. The seeding efficiency was assessed with MC3T3-E1 individual cells and HepaRG™ spheroids. These data collapse into a single master curve that exhibits a sharp transition from 100 % to 0 %-efficiency as the entity diameter approaches the mean pore size in the dry state. Altogether, we can thus precisely tune the porosity of these 3D materials of interest for 3D cell culture and cGMP production for tissue engineering.

Trajectory of Pain, Functional Limitation, and Parental Coping Resources Following Pediatric Short-stay Surgery: Factors Impacting Rate of Recovery.

OBJECTIVES: Although there are many benefits of short-stay hospital admissions for high volume, pediatric surgical procedures, this model of care places greater responsibility on parents for the management of children's pain. This study aimed to document the trajectory of child pain outcomes and a range of parent-reported functional outcomes following discharge from a short-stay surgical admission. Moreover, we aimed to document the trajectory of parental perceived personal coping resources. Second, we assessed whether parental dispositional factors, assessed before hospital discharge, predicted the child's pain intensity and parent-reported functional recovery. METHODS: Participants included children (aged 4 to 14 y) admitted for a short-stay tonsillectomy or appendectomy, and their parents. Parents completed a questionnaire before discharge from hospital. Demographic and surgical information was recorded from medical records. Following discharge, daily assessments of pain and functioning were carried out over a 10-day period using iPods or mobile phones. Predischarge and postdischarge data were obtained for 55 child and parent dyads. RESULTS: Pain intensity scores returned to low levels (2/10 or less) by day 5 for appendectomy and day 10 for tonsillectomy. Parents' perceived personal coping resources increased more slowly following tonsillectomy than appendectomy. Controlling for time since surgery and parental coping resources, parental pain-related catastrophizing was a significant predictor of child pain and functional recovery. DISCUSSION: Short-stay surgery results in parents facing considerable burden in managing their child's pain and functional impairment over a 10-day period. The potential value of screening for parental pain-related catastrophizing before discharge from hospital warrants further consideration and may enable identification of children likely to experience poorer recovery.

Channeling Effect and Tissue Morphology in a Perfusion Bioreactor Imaged by X-Ray Microtomography.

BACKGROUND: Perfusion bioreactors for tissue engineering hold great promises. Indeed, the perfusion of culture medium enhances species transport and mechanically stimulates the cells, thereby increasing cell proliferation and tissue formation. Nonetheless, their development is still hampered by a lack of understanding of the relationship between mechanical cues and tissue growth. METHODS: Combining tissue engineering, three-dimensional visualization and numerical simulations, we analyze the morphological evolution of neo-tissue in a model bioreactor with respect to the local flow pattern. NIH-3T3 cells were grown under perfusion for one, two and three weeks on a stack of 2 mm polyacetal beads. The model bioreactor was then imaged by X-ray micro-tomography and local tissue morphology was analyzed. To relate experimental observations and mechanical stimulii, a computational fluid dynamics model of flow around spheres in a canal was developed and solved using the finite element method. RESULTS: We observe a preferential tissue formation at the bioreactor periphery, and relate it to a channeling effect leading to regions of higher flow intensity. Additionally, we find that circular crater-like tissue patterns form in narrow channel regions at early culture times. Using computational fluid dynamic simulations, we show that the location and morphology of these patterns match those of shear stress maxima. Finally, the morphology of the tissue is qualitatively described as the tissue grows and reorganizes itself. CONCLUSION: Altogether, our study points out the key role of local flow conditions on the tissue morphology developed on a stack of beads in perfusion bioreactors and provides new insights for effective design of hydrodynamic bioreactors for tissue engineering using bead packings.

Alginate-encapsulated HepG2 cells in a fluidized bed bioreactor maintain function in human liver failure plasma.

Alginate-encapsulated HepG2 cells cultured in microgravity have the potential to serve as the cellular component of a bioartificial liver. This study investigates their performance in normal and liver failure (LF) human plasma over 6-8 h in a fluidized bed bioreactor. After 8 days of microgravity culture, beads containing 1.5 x 10(9) cells were perfused for up to 8 h at 48 mL/min with 300 mL of plasma. After exposure to 90% LF plasma, vital dye staining showed maintained cell viability, while a 7% increase in lactate dehydrogenase activity indicated minimal cell damage. Glucose consumption, lactate production, and a 4.3-fold linear increase in alpha-fetoprotein levels were observed. Detoxificatory function was demonstrated by quantification of bilirubin conjugation, urea synthesis, and Cyp450 1A activity. These data show that in LF plasma, alginate-encapsulated HepG2 cells can maintain viability, and metabolic, synthetic, and detoxificatory activities, indicating that the system can be scaled-up to form the biological component of a bioartificial liver.

Histological Method to Study the Effect of Shear Stress on Cell Proliferation and Tissue Morphology in a Bioreactor.

Background: Tissue engineering represents a promising approach for the production of bone substitutes. The use of perfusion bioreactors for the culture of bone-forming cells on a three-dimensional porous scaffold resolves mass transport limitations and provides mechanical stimuli. Despite the recent and important development of bioreactors for tissue engineering, the underlying mechanisms leading to the production of bone substitutes remain poorly understood. Methods: In order to study cell proliferation in a perfusion bioreactor, we propose a simplified experimental set-up using an impermeable scaffold model made of 2 mm diameter glass beads on which mechanosensitive cells, NIH-3T3 fibroblasts are cultured for up to 3 weeks under 10 mL/min culture medium flow. A methodology combining histological procedure, image analysis and analytical calculations allows the description and quantification of cell proliferation and tissue production in relation to the mean wall shear stress within the bioreactor. Results: Results show a massive expansion of the cell phase after 3 weeks in bioreactor compared to static control. A scenario of cell proliferation within the three-dimensional bioreactor porosity over the 3 weeks of culture is proposed pointing out the essential role of the contact points between adjacent beads. Calculations indicate that the mean wall shear stress experienced by the cells changes with culture time, from about 50 mPa at the beginning of the experiment to about 100 mPa after 3 weeks. Conclusion: We anticipate that our results will help the development and calibration of predictive models, which rely on estimates and morphological description of cell proliferation under shear stress.

Mechanisms of pore formation in hydrogel scaffolds textured by freeze-drying.

Whereas freeze-drying is a widely used method to produce porous hydrogel scaffolds, the mechanisms of pore formation involved in this process remained poorly characterized. To explore this, we focused on a cross-linked polysaccharide-based hydrogel developed for bone tissue engineering. Scaffolds were first swollen in 0.025% NaCl then freeze-dried at low cooling rate, i.e. -0.1 °C min-1, and finally swollen in aqueous solvents of increasing ionic strength. We found that scaffold's porous structure is strongly conditioned by the nucleation of ice. Electron cryo-microscopy of frozen scaffolds demonstrates that each pore results from the growth of one to a few ice grains. Most crystals were formed by secondary nucleation since very few nucleating sites were initially present in each scaffold (0.1 nuclei cm-3 °C-1). The polymer chains are rejected in the intergranular space and form a macro-network. Its characteristic length scale coincides with the ice grain size (160 µm) and is several orders of magnitude greater than the mesh size (90 nm) of the cross-linked network. After sublimation, the ice grains are replaced by macro-pores of 280 µm mean size and the resulting dry structure is highly porous, i.e. 93%, as measured by high-resolution X-ray tomography. In the swollen state, the scaffold mean pore size decreases in aqueous solvent of increasing ionic strength (120 µm in 0.025% NaCl and 54 µm in DBPS) but the porosity remains the same, i.e. 29% regardless of the solvent. Finally, cell seeding of dried scaffolds demonstrates that the pores are adequately interconnected to allow homogenous cell distribution. STATEMENT OF SIGNIFICANCE: The fabrication of hydrogel scaffolds is an important research area in tissue engineering. Hydrogels are textured to provide a 3D-framework that is favorable for cell proliferation and/or differentiation. Optimum hydrogel pore size depends on its biological application. Producing porous hydrogels is commonly achieved through freeze-drying. However, the mechanisms of pore formation remain to be fully understood. We carefully analyzed scaffolds of a cross-linked polysaccharide-based hydrogel developed for bone tissue engineering, using state-of-the-art microscopic techniques. Our experimental results evidenced the shaping of hydrogel during the freezing step, through a specific ice-templating mechanism. These findings will guide the strategies for controlling the porous structure of hydrogel scaffolds.

Assessment of the interplay between scaffold geometry, induced shear stresses, and cell proliferation within a packed bed perfusion bioreactor.

By favoring cell proliferation and differentiation, perfusion bioreactors proved efficient at optimizing cell culture. The aim of this study was to quantify cell proliferation within a perfusion bioreactor and correlate it to the wall shear stress (WSS) distribution by combining 3-D imaging and computational fluid dynamics simulations.NIH-3T3 fibroblasts were cultured onto a scaffold model made of impermeable polyacetal spheres or Polydimethylsiloxane cubes. After 1, 2, and 3 weeks of culture, constructs were analyzed by micro-computed tomography (µCT) and quantification of cell proliferation was assessed. After 3 weeks, the volume of cells was found four times higher in the stacking of spheres than in the stacking of cube.3D-µCT reconstruction of bioreactors was used as input for the numerical simulations. Using a lattice-Boltzmann method, we simulated the fluid flow within the bioreactors. We retrieved the WSS distribution (PDF) on the scaffolds surface at the beginning of cultivation and correlated this distribution to the local presence of cells after 3 weeks of cultivation. We found that the WSS distributions strongly differ between spheres and cubes even if the porosity and the specific wetted area of the stackings were very similar. The PDF is narrower and the mean WSS is lower for cubes (11 mPa) than for spheres (20 mPa). For the stacking of spheres, the relative occupancy of the surface sites by cells is maximal when WSS is greater than 20 mPa. For cubes, the relative occupancy is maximal when the WSS is lower than 10 mPa. The discrepancies between spheres and cubes are attributed to the more numerous sites in stacking of spheres that may induce 3-D (multi-layered) proliferation.

Mesoscale porosity at the dentin-enamel junction could affect the biomechanical properties of teeth.

In this paper, the 3D-morphology of the porosity in dentin is investigated within the first 350µm from the dentin-enamel junction (DEJ) by fluorescence confocal laser scanning microscopy (CLSM). We found that the porous microstructure exhibits a much more complex geometry than classically described, which may impact our fundamental understanding of the mechanical behavior of teeth and could have practical consequences for dental surgery. Our 3D observations reveal numerous fine branches stemming from the tubules which may play a role in cellular communication or mechanosensing during the early stages of dentinogenesis. The effect of this highly branched microstructure on the local mechanical properties is investigated by means of numerical simulations. Under simplified assumptions on the surrounding tissue characteristics, we find that the presence of fine branches negatively affects the mechanical properties by creating local stress concentrations. However, this effect is reduced by the presence of peritubular dentin surrounding the tubules. The porosity was also quantified using the CSLM data and compared to this derived from SEM imaging. A bimodal distribution of channel diameters was found near the DEJ with a mean value of 1.5-2µm for the tubules and 0.3-0.5µm for the fine branches which contribute to 30% of the total porosity (∼1.2%). A gradient in the branching density was observed from the DEJ towards the pulp, independently of the anatomical location. Our work constitutes an incentive towards more elaborate multiscale studies of dentin microstructure to better assess the effect of aging and for the design of biomaterials used in dentistry, e.g. to ensure more efficient bonding to dentin. Finally, our analysis of the tubular network structure provides valuable data to improve current numerical models.

Averaged model for momentum and dispersion in hierarchical porous media.

Hierarchical porous media are multiscale systems, where different characteristic pore sizes and structures are encountered at each scale. Focusing the analysis to three pore scales, an upscaling procedure based on the volume-averaging method is applied twice, in order to obtain a macroscopic model for momentum and diffusion-dispersion. The effective transport properties at the macroscopic scale (permeability and dispersion tensors) are found to be explicitly dependent on the mesoscopic ones. Closure problems associated to these averaged properties are numerically solved at the different scales for two types of bidisperse porous media. Results show a strong influence of the lower-scale porous structures and flow intensity on the macroscopic effective transport properties.

The inhibition of cell spreading on a cellulose substrate (cuprophan) induces an apoptotic process via a mitochondria-dependent pathway.

Cell shape was found to be a strong indicator of whether individual cells grow or die, and may play an important role in controlling apoptosis as well as cell growth. We compared here the behaviour of rounded Swiss 3T3 cells aggregated on a cellulose cuprophan membrane to those cultured on dish polystyrene. We demonstrated that cells aggregated on cellulose substrates for up to 48 h underwent programmed cell death that was associated with phosphatidylserine flipping and caspase 9 and caspase 3 activation, suggesting a mitochondria-dependent apoptotic process. In addition, we found that this phenomenon cannot be entirely explained by disengagement of alpha 5 beta 1 integrin ligation.

In vitro assessment of encapsulated C3A hepatocytes functions in a fluidized bed bioreactor.

In the present in vitro model, the authors intended to assess viability and functionality of hepatocytes encapsulated into alginate beads and submitted to a fluidized bed motion in a bioreactor. Human immortalized C3A line was chosen as cell model. Two controls consisting of (1) cells cultured on flasks and (2) cells encapsulated in alginate beads under static conditions were implemented. The cell functions studied were total protein, albumin, urea, and ammonia synthesis, as well as ammonia removal in the case of overdose. The comparison among the three cases studied showed that the three-dimensional structure of alginate offered a suitable environment for cell functions. In addition, the fluidized bed bioreactor enhanced the mass transfer and thus increased the amount of species released out of the beads, as compared with the static case. Ammonia detoxification only appeared reduced by encapsulation. The concept of a fluidized bed bioartificial liver was thus validated by this in vitro model, which indicated that cell functions could be efficiently retained. In addition, as far as urea and protein synthesis and release were concerned, the use of the C3A cell line, in combination with encapsulation and fluidization technology, offered a real potentiality for the purpose of extracorporeal liver supply.

Multiclass feature selection with kernel Gram-matrix-based criteria.

Feature selection has been an important issue in recent decades to determine the most relevant features according to a given classification problem. Numerous methods have emerged that take into account support vector machines (SVMs) in the selection process. Such approaches are powerful but often complex and costly. In this paper, we propose new feature selection methods based on two criteria designed for the optimization of SVM: kernel target alignment and kernel class separability. We demonstrate how these two measures, when fully expressed, can build efficient and simple methods, easily applicable to multiclass problems and iteratively computable with minimal memory requirements. An extensive experimental study is conducted both on artificial and real-world datasets to compare the proposed methods to state-of-the-art feature selection algorithms. The results demonstrate the relevance of the proposed methods both in terms of performance and computational cost.

A perfusion bioreactor for engineering bone constructs: an in vitro and in vivo study.

A perfusion bioreactor, which was designed based on fluidized bed concepts, was validated for the culture of bone constructs of clinically relevant size. For this study, natural coral has been used as three-dimensional scaffolds. This biomaterial is a microporous, biocompatible, osteoconductive, and absorbable scaffold. This perfusion bioreactor provided a stable environment in terms of osmolarity, pH, and, most importantly, oxidative stress. Bone constructs engineered in this system resulted in significantly higher cell proliferation and homogenous cell distribution than those cultured under static conditions. Particularly relevant to the production of bioengineered bone in a clinical setting, custom-made bone constructs (each one with volume up to 30 cm(3)) could be produced using a such perfusion bioreactor. Last, but not least, the bone constructs of clinically relevant volume thus produced were shown to be osteogenic when transplanted subcutaneously in sheep.