Project description:Reference point indentation (RPI) is a microindentation technique involving 20 cycles of loading in "force-control" that can directly assess a patient׳s bone tissue properties. Even though preliminary clinical studies indicate a capability for fracture discrimination, little is known about what mechanical behavior the various RPI properties characterize and how these properties relate to traditional mechanical properties of bone. To address this, the present study investigated the sensitivity of RPI properties to anatomical location and tissue organization as well as examined to what extent RPI measurements explain the intrinsic mechanical properties of human cortical bone. Multiple indents with a target force of 10N were done in 2 orthogonal directions (longitudinal and transverse) per quadrant (anterior, medial, posterior, and lateral) of the femoral mid-shaft acquired from 26 donors (25-101 years old). Additional RPI measurements were acquired for 3 orthogonal directions (medial only). Independent of age, most RPI properties did not vary among these locations, but they did exhibit transverse isotropy such that resistance to indentation is greater in the longitudinal (axial) direction than in the transverse direction (radial or circumferential). Next, beam specimens (~2mm×5mm×40mm) were extracted from the medial cortex of femoral mid-shafts, acquired from 34 donors (21-99 years old). After monotonically loading the specimens in three-point bending to failure, RPI properties were acquired from an adjacent region outside the span. Indent direction was orthogonal to the bending axis. A significant inverse relationship was found between resistance to indentation and the apparent-level mechanical properties. Indentation distance increase (IDI) and a linear combination of IDI and the loading slope, averaged over cycles 3 through 20, provided the best explanation of the variance in ultimate stress (r(2)=0.25, p=0.003) and toughness (r(2)=0.35, p=0.004), respectively. With a transverse isotropic behavior akin to tissue hardness and modulus as determined by micro- and nano-indentation and a significant association with toughness, RPI properties are likely influenced by both elastic and plastic behavior of bone tissue.
Project description:We have employed DNA methylation analysis using resected brain tissues from patients with epilepsy to investigate the pathophysiology of post-operative delirium (POD). POD is related to high mortality and long-term cognitive failure, but its mechanism is still unclear. This study represents the first transcriptome analysis of POD brain and provide new insight into POD pathology and prediction.
Project description:The brain is a complex organ made up of many different functional and structural regions consisting of different types of cells such as neurons and glia, as well as complex anatomical geometries. It is hypothesized that the different regions of the brain exhibit significantly different mechanical properties, which may be attributed to the diversity of cells and anisotropy of neuronal fibers within individual brain regions. The regional dynamic mechanical properties of P56 mouse brain tissue in vitro and in situ at velocities of 0.71-4.28 mm/s, up to a deformation of 70 μm are presented and discussed in the context of traumatic brain injury. The experimental data obtained from micro-indentation measurements were fit to three hyperelastic material models using the inverse Finite Element method. The cerebral cortex elicited a stiffer response than the cerebellum, thalamus, and medulla oblongata regions for all velocities. The thalamus was found to be the least sensitive to changes in velocity, and the medulla oblongata was most compliant. The results show that different regions of the mouse brain possess significantly different mechanical properties, and a significant difference also exists between the in vitro and in situ brain.
Project description:We have employed a single nucleous sequencing approach using 10x Genomics platform to investigate the pathophysiology of post-operative delirium (POD). POD is related to high mortality and long-term cognitive failure, but its mechanism is still unclear. This study represents the first transcriptome analysis of POD brain and provide new insight into POD pathology and prediction.
Project description:In vitro, animal cells are mostly cultured on a gel substrate. It was recently shown that substrate stiffness affects cellular behaviors in a significant way, including adhesion, differentiation, and migration. Therefore, an accurate method is needed to characterize the modulus of the substrate. In situ microscopic measurements of the gel substrate modulus are based on Hertz contact mechanics, where Young's modulus is derived from the indentation force and displacement measurements. In Hertz theory, the substrate is modeled as a linear elastic half-space with an infinite depth, whereas in practice, the thickness of the substrate, h, can be comparable to the contact radius and other relevant dimensions such as the radius of the indenter or steel ball, R. As a result, measurements based on Hertz theory overestimate the Young's modulus. In this work, we discuss the limitations of Hertz theory and then modify it, taking into consideration the nonlinearity of the material and large deformation using a finite-element method. We present our results in a simple correction factor, ψ, the ratio of the corrected Young's modulus and the Hertz modulus in the parameter regime of δ/h ≤ min (0.6, R/h) and 0.3 ≤R/h ≤ 12.7. The ψ factor depends on two dimensionless parameters, R/h and δ/h (where δ is the indentation depth), both of which are easily accessible to experiments. This correction factor agrees with experimental observations obtained with the use of polyacrylamide gel and a microsphere indentation method in the parameter range of 0.1 ≤δ/h ≤ 0.4 and 0.3 ≤R/h ≤ 6.2. The effect of adhesion on the use of Hertz theory for small indentation depth is also discussed.
Project description:The linker of nucleoskeleton and cytoskeleton (LINC) complex is responsible for tethering the nucleus to the cytoskeleton, providing a pathway for the cell's nucleus to sense mechanical signals from the environment. Recently, we explored the role of the LINC complex in the development of glandular epithelial acini, such as those found in kidneys, breasts, and other organs. Acini developed with disrupted LINC complexes exhibited a loss of structural integrity, including filling of the lumen structures. As part of our investigation, we performed a mechanical indentation assay of LINC disrupted and undisrupted MDCK II cells using a micro-indentation instrument mounted above a laser-scanning confocal microscope. Through a combination of force measurements acquired from the micro-indentation instrument and contact area measurements taken from fluorescence images, we determined the average contact pressure at which the acini structure ruptured. Here, we provide a detailed description of the design of the micro-indentation instrument, as well as the experimental steps developed to perform these bio-indentation measurements. Furthermore, we discuss the data analysis steps necessary to determine the rupture pressure of the acini structures. While this protocol is focused on the indentation of individual glandular acini, the methods presented here can be adapted to perform a variety of mechanical indentation experiments for both 2D and 3D biological systems.
Project description:Brain injuries are often characterized by diffusely distributed axonal and vascular damage invisible to medical imaging techniques. The spatial distribution of mechanical stresses and strains plays an important role, but is not sufficient to explain the diffuse distribution of brain lesions. It remains unclear how forces are transferred from the organ to the cell scale and why some cells are damaged while neighboring cells remain unaffected. To address this knowledge gap, we subjected histologically stained fresh human and porcine brain tissue specimens to compressive loading and simultaneously tracked cell and blood vessel displacements. Our experiments reveal different mechanisms of load transfer from the organ or tissue scale to single cells, axons, and blood vessels. Our results show that cell displacement fields are inhomogeneous at the interface between gray and white matter and in the vicinity of blood vessels-locally inducing significant deformations of individual cells. These insights have important implications to better understand injury mechanisms and highlight the importance of blood vessels for the local deformation of the brain's cellular structure during loading.
Project description:The mechanical function of the uterus is critical for a successful pregnancy. During gestation, uterine tissue grows and stretches to many times its size to accommodate the growing fetus, and it is hypothesized the magnitude of uterine tissue stretch triggers the onset of contractions. To establish rigorous mechanical testing protocols for the human uterus in hopes of predicting tissue stretch during pregnancy, this study measures the anisotropic mechanical properties of the human uterus using optical coherence tomography (OCT), instrumented spherical indentation, and video extensometry. In this work, we perform spherical indentation and digital image correlation to obtain the tissue's force and deformation response to a ramp-hold loading regimen. We translate previously reported fiber architecture, measured via optical coherence tomography, into a constitutive fiber composite material model to describe the equilibrium material behavior during indentation. We use an inverse finite element method integrated with a genetic algorithm (GA) to fit the material model to our experimental data. We report the mechanical properties of human uterine specimens taken across different anatomical locations and layers from one non-pregnant (NP) and one pregnant (PG) patient; both patients had pathological uterine tissue. Compared to NP uterine tissue, PG tissue has a more dispersed fiber distribution and equivalent stiffness material parameters. In both PG and NP uterine tissue, the mechanical properties differ significantly between anatomical locations.
Project description:The indentation test is widely used to determine the in situ biomechanical properties of articular cartilage. The mechanical parameters estimated from the test depend on the constitutive model adopted to analyze the data. Similar to most connective tissues, the solid matrix of cartilage displays different mechanical properties under tension and compression, termed tension-compression nonlinearity (TCN). In this study, cartilage was modeled as a porous elastic material with either a conewise linear elastic matrix with cubic symmetry or a solid matrix reinforced by a continuous fiber distribution. Both models are commonly used to describe the TCN of cartilage. The roles of each mechanical property in determining the indentation response of cartilage were identified by finite element simulation. Under constant loading, the equilibrium deformation of cartilage is mainly dependent on the compressive modulus, while the initial transient creep behavior is largely regulated by the tensile stiffness. More importantly, altering the permeability does not change the shape of the indentation creep curves, but introduces a parallel shift along the horizontal direction on a logarithmic time scale. Based on these findings, a highly efficient curve-fitting algorithm was designed, which can uniquely determine the three major mechanical properties of cartilage (compressive modulus, tensile modulus, and permeability) from a single indentation test. The new technique was tested on adult bovine knee cartilage and compared with results from the classic biphasic linear elastic curve-fitting program.
Project description:There is a strong demand for nanoindentation methods to probe the heterogeneous viscoelastic properties of soft tissues. Important applications include diagnosis of early onset diseases such as arthritis and investigations into cellular mechanoresponse in tissue. Quantification of tissue mechanics at length and time scales relevant to biological processes, however, remains a technical challenge. Here, we present a new nanoindentation approach that is ideally suited to probe the viscoelastic properties of soft, hydrated tissues. We built a ferrule-top probe that uses wavelength modulation in a Fabry-Pérot cavity configuration to detect cantilever deflection and to drive a feedback-controlled piezoelectric actuator. This technique allows us to control the static load applied onto the sample using an all-optical mm-sized probe. We extract the local elastic and viscous moduli of the samples by superposing a small oscillatory load and recording the indentation depth at the frequency of oscillation. By using a set of silicone elastomers with a range of stiffnesses representative of biological tissues, we demonstrate that the technique can accurately determine moduli over a wide range (0.1-100 kPa) and over a frequency range of 0.01-10 Hz. Direct comparison with macroscopic rheology measurements yields excellent quantitative agreement, without any fitting parameters. Finally, we show how this method can provide a spatially-resolved map of large variations in mechanical properties (orders of magnitude) across the surface of soft samples thanks to high sensitivity over large (>μm) cantilever deflections. This approach paves the way to investigations into the local dynamic mechanical properties of biological soft matter.