A Traumatic Impact Immediately Changes the Mechanical Properties of Articular Cartilage.

OBJECTIVE: To investigate whether and how a single traumatic impact changes the mechanical properties of talar articular cartilage. DESIGN: A marble was placed on the joint surface and a weight was dropped on both medial and lateral caprine talus to create a well-defined single focal impact. The mechanical properties of intact and impacted talar cartilage were measured with a micro-indenter. Elastic (storage) and viscous (loss) moduli were determined by oscillatory ramp and dynamic mechanical analysis protocols. RESULTS: We found significant differences between ankles and within the same ankle joint, with the medial talus having significantly higher storage- and loss moduli than the lateral talus. The storage- and loss moduli of intact articular cartilage increased with greater indentation depths. However, postimpact the storage- and loss moduli were significantly and consistently lower in all specimens indicating immediate posttraumatic damage. The deeper regions of talar cartilage were less affected by the impact than the more superficial regions. CONCLUSIONS: A single traumatic impact results in an immediate and significant decrease of storage- and loss moduli. Further research must focus on the development of non- or minimally invasive diagnostic tools to address the exact microdamage caused by the impact. We speculate that the traumatic impact damaged the collagen fibers that confine the water-binding proteoglycans and thereby decreasing the hydrostatic pressure of cartilage. As part of the treatment directly after a trauma, one could imagine a reduction or restriction of peak loads to prevent the progression of the cascade towards PTOA of the ankle joint.

Mechanical alterations of the hippocampus in the APP/PS1 Alzheimer's disease mouse model.

There is increasing evidence of altered tissue mechanics in neurodegeneration. However, due to difficulties in mechanical testing procedures and the complexity of the brain, there is still little consensus on the role of mechanics in the onset and progression of neurodegenerative diseases. In the case of Alzheimer's disease (AD), magnetic resonance elastography (MRE) studies have indicated viscoelastic differences in the brain tissue of AD patients and healthy controls. However, there is a lack of viscoelastic data from contact mechanical testing at higher spatial resolution. Therefore, we report viscoelastic maps of the hippocampus obtained by a dynamic indentation on brain slices from the APP/PS1 mouse model where individual brain regions are resolved. A comparison of viscoelastic parameters shows that regions in the hippocampus of the APP/PS1 mice are significantly stiffer than wild-type (WT) mice and have increased viscous dissipation. Furthermore, indentation mapping at the cellular scale directly on the plaques and their surroundings did not show local alterations in stiffness although overall mechanical heterogeneity of the tissue was high (SD∼40%).

Viscoelastic properties of white and gray matter-derived microglia differentiate upon treatment with lipopolysaccharide but not upon treatment with myelin.

BACKGROUND: The biomechanical properties of the brain have increasingly been shown to relate to brain pathology in neurological diseases, including multiple sclerosis (MS). Inflammation and demyelination in MS induce significant changes in brain stiffness which can be linked to the relative abundance of glial cells in lesions. We hypothesize that the biomechanical, in addition to biochemical, properties of white (WM) and gray matter (GM)-derived microglia may contribute to the differential microglial phenotypes as seen in MS WM and GM lesions. METHODS: Primary glial cultures from WM or GM of rat adult brains were treated with either lipopolysaccharide (LPS), myelin, or myelin+LPS for 24 h or left untreated as a control. After treatment, microglial cells were indented using dynamic indentation to determine the storage and loss moduli reflecting cell elasticity and cell viscosity, respectively, and subsequently fixed for immunocytochemical analysis. In parallel, gene expression of inflammatory-related genes were measured using semi-quantitative RT-PCR. Finally, phagocytosis of myelin was determined as well as F-actin visualized to study the cytoskeletal changes. RESULTS: WM-derived microglia were significantly more elastic and more viscous than microglia derived from GM. This heterogeneity in microglia biomechanical properties was also apparent when treated with LPS when WM-derived microglia decreased cell elasticity and viscosity, and GM-derived microglia increased elasticity and viscosity. The increase in elasticity and viscosity observed in GM-derived microglia was accompanied by an increase in Tnfα mRNA and reorganization of F-actin which was absent in WM-derived microglia. In contrast, when treated with myelin, both WM- and GM-derived microglia phagocytose myelin decrease their elasticity and viscosity. CONCLUSIONS: In demyelinating conditions, when myelin debris is phagocytized, as in MS lesions, it is likely that the observed differences in WM- versus GM-derived microglia biomechanics are mainly due to a difference in response to inflammation, rather than to the event of demyelination itself. Thus, the differential biomechanical properties of WM and GM microglia may add to their differential biochemical properties which depend on inflammation present in WM and GM lesions of MS patients.

Indentation probe with optical fibre array-based optical coherence tomography for material deformation.

We present a new optomechanical probe for mechanical testing of soft matter. The probe consists of a micromachined cantilever equipped with an indenting sphere, and an array of 16 single-mode optical fibres, which are connected to an optical coherence tomography (OCT) system that allows subsurface analysis of the sample during the indentation stroke. To test our device and its capability, we performed indentation on a PDMS-based phantom. Our findings demonstrate that Common Path (CP)-OCT via lensed optical fibres can be successfully combined with a microindentation sensor to visualise the phantom's deformation profile at different indentation depths and locations in real time. LAY DESCRIPTION: This work presents a new approach to simultaneously perform micro-indentation experiments and OCT imaging. An optical fiber array-based sensor is used to develop a new hybrid tool where micro-indentation is combined with optical coherence tomography. The sensor is therefore capable of compressing a sample with a small force and simultaneously collecting OCT depth profiles underneath and around the indentation point. This method offers the opportunity to characterize the mechanical properties of soft materials and simultaneously visualize their deformation profile. The ability to integrate OCT imaging with indentation technology is promising for the non-invasive and precise characterization of different soft materials.

Viscoelastic mapping of mouse brain tissue: Relation to structure and age.

There is growing evidence that mechanical factors affect brain functioning. However, brain components responsible for regulating the physiological mechanical environment are not completely understood. To determine the relationship between structure and stiffness of brain tissue, we performed high-resolution viscoelastic mapping by dynamic indentation of the hippocampus and the cerebellum of juvenile mice brains, and quantified relative area covered by neurons (NeuN-staining), axons (neurofilament NN18-staining), astrocytes (GFAP-staining), myelin (MBP-staining) and nuclei (Hoechst-staining) of juvenile and adult mouse brain slices. Results show that brain subregions have distinct viscoelastic parameters. In gray matter (GM) regions, the storage modulus correlates negatively with the relative area of nuclei and neurons, and positively with astrocytes. The storage modulus also correlates negatively with the relative area of myelin and axons (high cell density regions are excluded). Furthermore, adult brain regions are ∼ 20%-150% stiffer than the comparable juvenile regions which coincide with increase in astrocyte GFAP-staining. Several linear regression models are examined to predict the mechanical properties of the brain tissue based on (immuno)histochemical stainings.

Toward clinical elastography of dermal tissues: A medical device to probe skin's elasticity through suction, with subsurface imaging via optical coherence tomography.

The mechanical behavior of dermal tissues is unarguably recognized for its diagnostic ability and in the last decades received a steadily increasing interest in dermatology practices. Among the various methods to investigate the mechanics of skin in clinical environments, suction-based ones are especially noteworthy, thanks to their qualities of minimal invasiveness and relative simplicity of setups and data analysis. In such experiments, structural visualization of the sample is highly desirable, both in its own right and because it enables elastography. The latter is a technique that combines the knowledge of an applied mechanical stimulus and the visualization of the induced deformation to result in a spatially resolved map of the mechanical properties, which is particularly important for an inhomogeneous and layered material such as skin. We present a device, designed for clinical trials in dermatology practices, that uses a handheld probe to (1) deliver a suction-based, controlled mechanical stimulus and (2) visualize the subsurface structure via optical coherence tomography. We also present a device-agnostic data-analysis framework, consisting of a Python library, released in the public domain. We show the working principle of the setup on a polymeric model and on a volunteer's skin.

Dynamic indentation reveals differential viscoelastic properties of white matter versus gray matter-derived astrocytes upon treatment with lipopolysaccharide.

Astrocytes in white matter (WM) and gray matter (GM) brain regions have been reported to have different morphology and function. Previous single cell biomechanical studies have not differentiated between WM- and GM-derived samples. In this study, we explored the local viscoelastic properties of isolated astrocytes and show that astrocytes from rat brain WM-enriched areas are ~1.8 times softer than astrocytes from GM-enriched areas. Upon treatment with pro-inflammatory lipopolysaccharide, GM-derived astrocytes become significantly softer in the nuclear and the cytoplasmic regions, where the F-actin network appears rearranged, whereas WM-derived astrocytes preserve their initial mechanical features and show no alteration in the F-actin cytoskeletal network. We hypothesize that the flexibility in biomechanical properties of GM-derived astrocytes may contribute to promote regeneration of the brain under neuroinflammatory conditions.

Mapping the mechanical properties of paintings via nanoindentation: a new approach for cultural heritage studies.

A comprehensive understanding of the behaviour of the heterogenous layers within the paint stratigraphies in historical paintings is crucial to evaluate their long term stability. We aim to refine nanoindentation as a new tool to investigate the mechanical behaviour of historical oil paints, by adapting the probes and the protocol already used in biomechanical research on soft tissues. The depth-controlled indentation profile performed with a spherical probe provides an evaluation of the non-linear viscoelastic behaviour of the individual layers in paint at local scale. The technique is non-destructive and guarantees the integrity of the surface after indentation. The mapping of elasticity demonstrates the properties' heterogeneity of the composite material within the paint layers, as well as between the individual layers and their interfaces.

Photoacoustic spectroscopy for gas sensing: A comparison between piezoelectric and interferometric readout in custom quartz tuning forks.

We report on a comparison between piezoelectric and interferometric readouts of vibrations in quartz tuning forks (QTFs) when acting as sound wave transducers in a quartz-enhanced photoacoustic setup (QEPAS) for trace gas detection. A theoretical model relating the prong vibration amplitude with the QTF prong sizes and electrical resistance is proposed. To compare interferometric and piezoelectric readouts, two QTFs have been selected; a tuning fork with rectangular-shape of the prongs, having a resonance frequency of 3.4 kHz and a quality-factor of 4,000, and a QTF with prong having a T-shape characterized by a resonance frequency of 12.4 kHz with a quality-factor of 15,000. Comparison between the interferometric and piezoelectric readouts were performed by using both QTFs in a QEPAS sensor setup for water vapor detection. We demonstrated that the QTF geometry can be properly designed to enhance the signal from a specific readout mode.

In vivo characterization of chick embryo mesoderm by optical coherence tomography-assisted microindentation.

Embryos are growing organisms with highly heterogeneous properties in space and time. Understanding the mechanical properties is a crucial prerequisite for the investigation of morphogenesis. During the last 10 years, new techniques have been developed to evaluate the mechanical properties of biological tissues in vivo. To address this need, we employed a new instrument that, via the combination of micro-indentation with Optical Coherence Tomography (OCT), allows us to determine both, the spatial distribution of mechanical properties of chick embryos, and the structural changes in real-time. We report here the stiffness measurements on the live chicken embryo, from the mesenchymal tailbud to the epithelialized somites. The storage modulus of the mesoderm increases from (176 ± 18) Pa in the tail to (716 ± 117) Pa in the somitic region (mean ± SEM, n = 12). The midline has a mean storage modulus of (947 ± 111) Pa in the caudal (PSM) presomitic mesoderm (mean ± SEM, n = 12), indicating a stiff rod along the body axis, which thereby mechanically supports the surrounding tissue. The difference in stiffness between midline and presomitic mesoderm decreases as the mesoderm forms somites. This study provides an efficient method for the biomechanical characterization of soft biological tissues in vivo and shows that the mechanical properties strongly relate to different morphological features of the investigated regions.

Investigating the Effects of Mechanical Stimulation on Retinal Ganglion Cell Spontaneous Spiking Activity.

Mechanical forces are increasingly recognized as major regulators of several physiological processes at both the molecular and cellular level; therefore, a deep understanding of the sensing of these forces and their conversion into electrical signals are essential for studying the mechanosensitive properties of soft biological tissues. To contribute to this field, we present a dual-purpose device able to mechanically stimulate retinal tissue and to record the spiking activity of retinal ganglion cells (RGCs). This new instrument relies on combining ferrule-top micro-indentation, which provides local measurements of viscoelasticity, with high-density multi-electrode array (HD-MEAs) to simultaneously record the spontaneous activity of the retina. In this paper, we introduce this instrument, describe its technical characteristics, and present a proof-of-concept experiment that shows how RGC spiking activity of explanted mice retinas respond to mechanical micro-stimulations of their photoreceptor layer. The data suggest that, under specific conditions of indentation, the retina perceive the mechanical stimulation as modulation of the visual input, besides the longer time-scale of activation, and the increase in spiking activity is not only localized under the indentation probe, but it propagates across the retinal tissue.

Immersion photoacoustic spectrometer (iPAS) for arcing fault detection in power transformers.

We report on the development of the first immersion photoacoustic spectrometer (iPAS) for arcing fault detection in power transformers. The spectrometer consists of a detection system and an all-optical photoacoustic sensing head mounted inside a small permeable chamber where dissolved C2H2 diffuses while the transformer oil is kept out. Our all-optical iPAS sensor can be placed directly inside an oil bath and measure dissolved C2H2 with the sensitivity and linearity needed for in situ arcing fault detection. Moreover, its fast response time holds great promise for extra-early fault diagnosis.

Micro-indentation and optical coherence tomography for the mechanical characterization of embryos: Experimental setup and measurements on chicken embryos.

The investigation of the mechanical properties of embryos is expected to provide valuable information on the phenomenology of morphogenesis. It is thus believed that, by mapping the viscoelastic features of an embryo at different stages of growth, it may be possible to shed light on the role of mechanics in embryonic development. To contribute to this field, we present a new instrument that can determine spatiotemporal distributions of mechanical properties of embryos over a wide area and with unprecedented accuracy. The method relies on combining ferrule-top micro-indentation, which provides local measurements of viscoelasticity, with Optical Coherence Tomography, which can reveal changes in tissue morphology and help the user identify the indentation point. To prove the working principle, we have collected viscoelasticity maps of fixed and live HH11-HH12 chicken embryos. Our study shows that the instrument can reveal correlations between tissue morphology and mechanical behavior. STATEMENT OF SIGNIFICANCE: Local mechanical properties of soft biological tissue play a crucial role in several biological processes, including cell differentiation, cell migration, and body formation; therefore, measuring tissue properties at high resolution is of great interest in biology and tissue engineering. To provide an efficient method for the biomechanical characterization of soft biological tissues, we introduce a new tool in which the combination of non-invasive Optical Coherence Tomography imaging and depth-controlled indentation measurements allows one to map the viscoelastic properties of biological tissue and investigate correlations between local mechanical features and tissue morphology with unprecedented resolution.

A fiber-tip photoacoustic sensor for in situ trace gas detection.

Most trace gas detection methods developed so far largely rely on active sampling procedures, which are known to introduce different kinds of artifacts. Here, we demonstrate sampling-free in situ trace gas detection in millimeter scale volumes with fiber coupled cantilever enhanced photoacoustic spectroscopy. Our 2.4 mm diameter fiber-tip sensor is free from the wavelength modulation induced background signal (a phenomenon that is often overlooked in photoacoustic spectroscopy) and reaches a normalized noise equivalent absorption coefficient of 1.3 × 10-9 W cm-1 Hz-1/2 for acetylene detection. To validate its in situ gas detection capability, we inserted the sensor into a mini fermenter for headspace monitoring of CO2 production during yeast fermentation. Our results show that the sensor can easily follow the different stages of the CO2 production of the fermentation process in great detail.

70 µm diameter optical probe for common-path optical coherence tomography in air and liquids.

We investigate and validate a novel method to fabricate ultrathin optical probes for common-path optical coherence tomography (CP-OCT). The probes are obtained using a 65 µm barium titanate microsphere inserted into an inward concave cone chemically etched at the end of a single-mode fiber. We demonstrate that the high refractive index (n=1.95) of the barium titanate microspheres allows one to maintain high sensitivity even while imaging in liquids, reaching a sensitivity of 83 dB. Due to its low cost, flexibility, and ease of use, the probe holds promise for the development of a new generation of ultrathin needle-based OCT systems.

Comparison of frequency and strain-rate domain mechanical characterization.

Indentation is becoming increasingly popular to test soft tissues and (bio)materials. Each material exhibits an unknown intrinsic "mechanical behaviour". However, limited consensus on its "mechanical properties" (i.e. quantitative descriptors of mechanical behaviour) is generally present in the literature due to a number of factors, which include sample preparation, testing method and analysis model chosen. Viscoelastic characterisation - critical in applications subjected to dynamic loading conditions - can be performed in either the time- or frequency-domain. It is thus important to selectively investigate whether the testing domain affects the mechanical results or not. We recently presented an optomechanical indentation tool which enables both strain-rate (nano-[Formula: see text]) and frequency domain (DMA) measurements while keeping the sample under the same physical conditions and eliminating any other variability factor. In this study, a poly(dimethylsiloxane) sample was characterised with our system. The DMA data were inverted to the time-domain through integral transformations and then directly related to nano-[Formula: see text] strain-rate dependent results, showing that, even though the data do not perfectly overlap, there is an excellent correlation between them. This approach indicates that one can convert an oscillatory measurement into a strain-rate one and still capture the trend of the "mechanical behaviour" of the sample investigated.

Regional variations in stiffness in live mouse brain tissue determined by depth-controlled indentation mapping.

The mechanical properties of brain tissue play a pivotal role in neurodevelopment and neurological disorders. Yet, at present, there is no consensus on how the different structural parts of the tissue contribute to its stiffness variations. Here, we have gathered depth-controlled indentation viscoelasticity maps of the hippocampus of acute horizontal live mouse brain slices. Our results confirm the highly viscoelestic nature of brain tissue. We further show that the mechanical properties are non-uniform and at least related to differences in morphological composition. Interestingly, areas with higher nuclear density appear to be softer than areas with lower nuclear density.

Publisher Correction: Minimally Invasive Micro-Indentation: mapping tissue mechanics at the tip of an 18G needle.

A correction to this article has been published and is linked from the HTML and PDF versions of this paper. The error has not been fixed in the paper.

Stiffening of the nucleus pulposus upon axial loading of the intervertebral disc: An experimental in situ study.

Mechanical loading is inherently related to the function and degeneration of the intervertebral disc. We present a series of experiments aimed at measuring the effect of a loading/unloading cycle of the intervertebral disc on the mechanical properties of the nucleus pulposus. The study relies on our new minimally invasive microindenter, which allows us to quantify the storage and loss moduli of the nucleus pulposus by inserting an optomechanical probe in an intact (resected) intervertebral disk through the annulus fibrosis via a small needle. Our results indicate that, under the influence of compressive loading, the nucleus pulposus exhibits a more solid-like behavior.

Minimally Invasive Micro-Indentation: mapping tissue mechanics at the tip of an 18G needle.

Experiments regarding the mechanical properties of soft tissues mostly rely on data collected on specimens that are extracted from their native environment. During the extraction and in the time period between the extraction and the completion of the measurements, however, the specimen may undergo structural changes which could generate unwanted artifacts. To further investigate the role of mechanics in physiology and possibly use it in clinical practices, it is thus of paramount importance to develop instruments that could measure the viscoelastic response of a tissue without necessarily excising it. Tantalized by this opportunity, we have designed a minimally invasive micro-indenter that is able to probe the mechanical response of soft tissues, in situ, via an 18G needle. Here, we discuss its working principle and validate its usability by mapping the viscoelastic properties of a complex, confined sample, namely, the nucleus pulposus of the intervertebral disc. Our findings show that the mechanical properties of a biological tissue in its local environment may be indeed different than those that one would measure after excision, and thus confirm that, to better understand the role of mechanics in life sciences, one should always perform minimally invasive measurements like those that we have here introduced.