Tissue Viscoelastic Properties Research Articles

Characterisation of the soft tissue viscous and elastic properties using ultrasound elastography and rheological models: validation and applications in plantar soft tissue assessment

Objective. The mechanical behaviour of soft tissue is influenced by its elastic and viscous characteristics. Therefore, the aim of this study was to develop a validated method to characterise the viscoelastic properties of soft tissues based on ultrasound elastography data. Approach. Plantar soft tissue was chosen as the tissue of interest, and gelatine-phantoms replicating its mechanical properties were manufactured for validation of the protocol. Both plantar soft tissue and the phantom were scanned using Reverberant shear wave ultrasound (US) elastography at 400–600 Hz. Shear wave speed was estimated using the US particle velocity data. The viscoelastic parameters were extracted by fitting the Young’s modulus as a function of frequency derived from the constitutive equations of the eight rheological models (four classic and their fractional-derivative versions) to the shear wave dispersion data. Furthermore, stress-time functions derived from the eight rheological models were fitted to the phantom stress-relaxation data. Main results. The viscoelastic parameters estimated using elastography data based on the fractional-derivative (FD) models, compared to the classic models, were closer to those quantified using the mechanical test. In addition, the FD-Maxwell and FD-Kelvin–Voigt models showed to more effectively replicate the viscoelastic behaviour of the plantar soft tissue with minimum number of model parameters ( = 0.72 for both models) . Hence the FD-KV and FD-Maxwell models can more effectively quantify the viscoelastic characteristics of the soft tissue compared to other models. Significance. In this study, a method for mechanical characterisation of the viscoelastic properties of soft tissue in ultrasound elastography was developed and fully validated. An investigation into the most valid rheological model and its applications in plantar soft tissue assessment were also presented. This proposed approach for the characterisation of viscous and elastic mechanical properties of soft tissue has implications in assessing the soft tissue function where those can be used as markers for diagnosis or prognosis of tissue status.

ANKLE LIGAMENT REPAIR FOR RESTORATION OF FUNCTION: EXPLORING THE APPLICATION OF DECELLULARIZED GRAFTS

Ligament integrity is directly associated with ankle stability. Nearly 40% of ankle sprains result in chronic ankle instability, affecting biomechanics and potentially causing osteoarthritis. Ligament replacement could restore stability and avoid this degenerative pathway, but a greater understanding of ankle ligament behaviour is required. Additionally, autograft or allograft use is limited by donor-site morbidity and inflammatory responses respectively. Decellularised porcine grafts could address this, by removing cellular material to prevent acute immune responses, while preserving mechanical properties.This project will characterise commonly injured ankle ligaments and damage mechanisms, identify ligament reconstruction requirements, and investigate the potential of decellularised porcine grafts as a replacement material.Several porcine tendons were evaluated to identify suitable candidates for decellularisation. The viscoelastic properties of native tissues were assessed using dynamic mechanical analysis (DMA), followed by ramp to ‘sub-rupture’ at 1% strain/s, and further DMA. Multiple samples (n=5) were taken along the graft to assess variation along the tendon.When identifying suitable porcine tendons, a lack of literature on human ankle ligaments was identified. Inconsistencies in measurement methods and properties reported makes comparison between studies difficult.Preliminary testing on porcine tendons suggested there is little variation in viscoelastic properties along the length of tendon. Testing also suggested strain rates of 1%/s sub-rupture was not large enough to affect viscoelastic properties (no changes in storage or loss moduli or tanẟ). Further testing is underway to improve upon low initial sample numbers and confirm these results, with varying strain rates to identify suitable sub-rupture sprain conditions.This work highlights need for new data on human ankle ligaments to address knowledge gaps and identify suitable replacement materials. Future work will generate this data and decellularise porcine tendons of similar dimensions. Collagen damage will be investigated using histology and lightsheet microscopy, and viscoelastic changes through DMA.

Characterizing viscoelastic properties of human melanoma tissue using Prony series.

Melanoma is the most invasive and deadly skin cancer, which causes most of the deaths from skin cancer. It has been demonstrated that the mechanical properties of tumor tissue are significantly altered. However, data about characterizing the mechanical properties of in vivo melanoma tissue are extremely scarce. In addition, the viscoelastic or viscous properties of melanoma tissue are rarely reported. In this study, we measured and quantitated the viscoelastic properties of human melanoma tissues based on the stress relaxation test, using the indentation-based mechanical analyzer that we developed previously. The melanoma tissues from eight patients of different ages (57-95), genders (male and female patients), races (White and Asian), and sites (nose, arm, shoulder, and chest) were excised and tested. The results showed that the elastic property (i.e., shear modulus) of melanoma tissue was elevated compared to normal tissue, while the viscous property (i.e., relaxation time) was reduced. Moreover, the tissue thickness had a significant impact on the viscoelastic properties, probably due to the amount of the adipose layer. Our findings provide new insights into the role of the viscous and elastic properties of melanoma cell mechanics, which may be implicated in the disease state and progression.

Data-Driven and Compliance-Based Fault-Tolerance for a Flexible and Extendable Robotic Implant Coupled to a Growing Tissue

Robotic implants for real-time and long-term monitoring and therapies are being researched and could open new frontiers in the medical field. For these devices to see widespread adoption, though, key challenges still need to be overcome, including reliability. Over the years, many computational techniques have been developed to impart fault-tolerance to robots and industrial plants. However, the application of these approaches to robotic implants is still challenging, due to the lack of information about the complex behavior of soft tissue (e.g. growth, viscoelasticity) and robot-tissue interaction. In this paper, a novel fault detection framework for a flexible extendable robotic implant is presented. Based on Canonical Correlation Analysis, the approach exploits the flexibility of the robot to extrapolate information for fault identification purposes. The experiments are conducted with a soft tissue simulator, which can emulate the viscoelastic properties of tissue as well as its growth, providing a realistic testing platform. The experiments prove the reliability of the flexible extendable robotic implant and its robustness to system-level external disturbances. Long-term tests are also presented, where the implant extends 80 mm, to its full length, counteracting simulated hardware faults over a 24-hour period, and provide a promising basis for future in-vivo trials.

Using uniaxial tensile testing to evaluate the biomechanical properties of bladder tissue after spinal cord injury in rat model

To investigate the biomechanical properties of rat bladder tissue after spinal cord injury (SCI) using uniaxial tensile testing.Evidence suggests the bladder wall undergoes remodeling following SCI. There is limited data describing the biomechanical properties of bladder wall after SCI. This study describes the changes in elastic and viscoelastic mechanical properties of bladder tissue using a rat model after SCI.Seventeen adult rats received mid-thoracic SCI. Basso, Beattie, and Bresnahan (BBB) locomotor testing was performed on the rats 7–14 days after injury quantifying the degree of SCI. Bladder tissue samples were collected from controls and spinal injured rats at 2- and 9-weeks post-injury. Tissue samples underwent uniaxial stress relaxation to determine instantaneous and relaxation modulus as well as monotonic load-to failure to determine Young’s modulus, yield stress and strain, and ultimate stress.SCI resulted in abnormal BBB locomotor scores. Nine weeks post-injury, instantaneous modulus decreased by 71.0% (p = 0.03) compared to controls. Yield strain showed no difference at 2 weeks post-injury but increased 78% (p = 0.003) in SCI rats at 9 weeks post-injury. Compared to controls, ultimate stress decreased 46.5% (p = 0.05) at 2 weeks post-injury in SCI rats but demonstrated no difference at 9 weeks post-injury.The biomechanical properties of rat bladder wall 2 weeks after SCI showed minimal difference compared to controls. By week 9, SCI bladders had a reduction in instantaneous modulus and increased yield strain. The findings indicate biomechanical differences can be identified between control and experimental groups at 2- and 9-week intervals using uniaxial testing.

A Microrobot With an Attached Microforce Sensor for Transurethral Access to the Bladder Interior Wall

AbstractWe present the conceptual design and limited functionality prototype and characterization of a system for application in transurethral palpation of any targeted area of the bladder interior wall tissue consisting of a robotic manipulator and a microforce sensor attached at its tip all less than 3.5 mm in diameter. A hyper-redundant ten-joint six degrees-of-freedom (6DOF) manipulator (5DOF rigid and five-joint continuum segments) is presented along with the forward and inverse kinematics analyses based on a Jacobian formulation to prevent configuration singularities. Simulated motion studies demonstrate the ability of the proposed manipulator to attain a desired pose (normal to the tissue) with any area in the bladder including the difficult to reach trigone area. A strain gauge-based microforce sensor is designed using finite element analysis (safety factor > 3), prototyped using additive manufacturing, and characterized. The characterized sensor was used to acquire in vivo measurements to evaluate human palm tissue viscoelastic properties. A single module of the continuum segment is designed and prototyped using additive manufacturing, and used to characterize its tension-bend angle behavior. Finite element analysis is used to improve structurally weak regions of the vertebra. A three-joint four-vertebrae prototype was successfully actuated to reach a bend state using tendons. The developed robot and sensor prototypes demonstrate capabilities of the proposed concept which could be a possible solution to quantitatively evaluate localized biomechanical properties of the bladder tissue to improve treatment and provide better patient care.

Nanoindentation study of the viscoelastic properties of human triple negative breast cancer tissues: Implications for mechanical biomarkers.

This paper presents the results of a combined experimental and theoretical study of the structure and viscoelastic properties of human non-tumorigenic mammary breast tissues and triple negative breast cancer (TNBC) tissues of different histological grades. A combination of immunofluorescence and confocal microscopy, and atomic force microscopy is used to study the actin cytoskeletal structures of non-tumorigenic and tumorigenic breast tissues (grade I to grade III). A combination of nanoindentation and statistical techniques is then used to measure viscoelastic properties of non-tumorigenic and human TNBC of different histological grades. A Standard Fluid Model/Anti-Zener Model II is also used to characterize the viscoelastic properties of the non-tumorigenic and tumorigenic TNBC tissues of different grades. The implications of the results are discussed for the potential application of nanoindentation and statistical deconvolution techniques to the development of mechanical biomarkers for TNBC detection/cancer diagnosis. STATEMENT OF SIGNIFICANCE: There is increasing interest in the development of mechanical biomarkers for cancer diagnosis. Here, we show that nanoindentation techniques can be used to characterize the viscoelastic properties of normal breast tissue and TNBC tissues of different histological grades. The Standard Fluid Model (Anti-Zener Model II) is used to classify the viscoelastic properties of breast tissues of different TNBC histological grades. Our results suggest that breast tissue and TNBC tissue viscoelastic properties can be used as mechanical biomarkers for the detection of TNBC at different stages.

Longitudinal motion of focused shear wave beams in soft elastic media.

Shear waves are employed in medical ultrasound imaging because they reveal variations in viscoelastic properties of soft tissue. Frequencies below 1 kHz are required due to the substantially higher attenuation and lower propagation speeds than for compressional waves. Shear waves exhibiting particle motion in the direction of propagation, referred to as longitudinal shear waves, can be generated with longitudinal motion of a circular disk on the surface of a soft elastic medium. This approach permits imaging of the longitudinal shear wave with a conventional ultrasound transducer that is coaxial with the source of the shear wave. Presented here is a mathematical model describing the complete wave field generated by displacement at the surface of an isotropic elastic half-space. Numerical simulations are shown for longitudinal, transverse, torsional, and radial source polarizations, with emphasis on focused longitudinal shear waves. Predictions are consistent with measurements of light beams revealing that the longitudinal electric field component produces a smaller focal spot than the transverse field component [Dorn, Quabis, and Leuchs, Phys. Rev. Lett. 91, 233901 (2003)]. Simulations are compared with preliminary measurements of a focused longitudinal shear wave beam generated in a soft tissue phantom by longitudinal motion of a spherically concave piston.

Injectable Double-Network Hydrogel-Based Three-Dimensional Cell Culture Systems for Regenerating Dental Pulp

The regeneration of dental pulp tissue is very important, but difficult, in dentistry. The biocompatibility, water content, and viscoelastic properties of pulp-like tissue must be optimized to achieve the efficient transfer of metabolites and nutrients, a suitable degradation rate, distribution of encapsulated cells, injectability, and gelation in situ under physiological conditions. As promising materials for pulp regeneration, hydrogel scaffolds have been produced to simulate the extracellular matrix and transmit signaling molecules. It is imperative to develop hydrogels to effectively regenerate pulp tissue for clinical application. Here, two injectable double-network (DN) hydrogel-based three-dimensional (3D) cell culture systems were developed for regenerating dental pulp. The microstructure, mechanical property, rheology property, and degradation behavior of the injectable DN glycol chitosan-based hydrogels in a simulated root canal model were characterized and compared to a single-network (SN) glycol chitosan-based hydrogel. Human dental pulp stem cells (hDPSCs) were then encapsulated into the GC-based hydrogels for the regeneration of pulp tissue, and the biological performance was investigated both in vitro and in vivo. The results showed that the DN hydrogels had ideal injectability under physiological conditions due to the dynamic nature of the crosslinks. Besides, the DN hydrogels exhibited better mechanical properties and longer degradation duration than the corresponding SN hydrogel. As a 3D cell culture system, the characteristics of the DN hydrogel facilitated odontogenic differentiation and mineralization of hDPSCs in vitro. Further in vivo analysis confirmed that the chemical composition, matrix stiffness, and degradation rate of the DN hydrogel matched those of pulp-like fibrous connective tissue, which might be related to Smad3 activation. These findings demonstrate that DN glycol chitosan-based hydrogels are suitable for the regeneration of pulp tissue.

Hippocampal subfield viscoelasticity in amnestic mild cognitive impairment evaluated with MR elastography

Hippocampal subfields (HCsf) are brain regions important for memory function that are vulnerable to decline with amnestic mild cognitive impairment (aMCI), which is often a preclinical stage of Alzheimer’s disease. Studies in aMCI patients often assess HCsf tissue integrity using measures of volume, which has little specificity to microstructure and pathology. We use magnetic resonance elastography (MRE) to examine the viscoelastic mechanical properties of HCsf tissue, which is related to structural integrity, and sensitively detect differences in older adults with aMCI compared to an age-matched control group. Group comparisons revealed HCsf viscoelasticity is differentially affected in aMCI, with CA1-CA2 and DG-CA3 exhibiting lower stiffness and CA1-CA2 exhibiting higher damping ratio, both indicating poorer tissue integrity in aMCI. Including HCsf stiffness in a logistic regression improves classification of aMCI beyond measures of volume alone. Additionally, lower DG-CA3 stiffness predicted aMCI status regardless of DG-CA3 volume. These findings showcase the benefit of using MRE in detecting subtle pathological tissue changes in individuals with aMCI via the HCsf particularly affected in the disease.

Dynamic changes in mechanical properties of the adult rat spinal cord after injury.

Spinal cord injury (SCI), a debilitating medical condition that can cause irreversible loss of neurons and permanent paralysis, currently has no cure. However, regenerative medicine may offer a promising treatment. Given that numerous regenerative strategies aim to deliver cells and materials in the form of tissue-engineered therapies, understanding and characterising the mechanical properties of the spinal cord tissue is very important. In this study, we have systematically characterised the spatiotemporal changes in elastic stiffness (elastic modulus, Pa) and viscosity (drop in peak force, %) of injured rat thoracic spinal cord tissues at distinct time points after crush injury using the indentation technique. Our results demonstrate that in comparison with uninjured spinal cord tissue, the injured tissues exhibited lower stiffness (median 3281 Pa versus 9632 Pa; P<0.001) but demonstrated elevated viscosity (median 80% versus 57%; P<0.001) at 3 days postinjury. Between 4 and 6 weeks after SCI, the overall viscoelastic properties of injured tissues returned to baseline values. At 12 weeks after SCI, in comparison with uninjured tissue, the injured spinal cord tissues displayed a significant increase in both elasticity (median 13698 Pa versus 9920 Pa; P<0.001) and viscosity (median 64% versus 58%; P<0.001). This work constitutes the first quantitative mapping of spatiotemporal changes in spinal cord tissue elasticity and viscosity in injured rats, providing a mechanical basis of the tissue for future studies on the development of biomaterials for SCI repair. STATEMENT OF SIGNIFICANCE: Spinal cord injury (SCI) is a devastating disease often leading to permanent paralysis. While enormous progress in understanding the molecular pathomechanisms of SCI has been made, the mechanical properties of injured spinal cord tissue have received considerably less attention. This study provides systematic characterization of the biomechanical evolution of rat spinal cord tissue after SCI using a microindentation test method. We find spinal cord tissue behaves significantly softer but more viscous immediately postinjury. As time passes, the lesion site gradually returns to baseline values and then displays pronounced increased viscoelastic properties. As host tissue mechanical properties are a crucial consideration for any biomaterial implanted into central nervous system, our results may have important implications for further studies of SCI repair.

Measurement of Force and Intramuscular Pressure Changes Related to Thrust Spinal Manipulation in an In Vivo Animal Model.

Current knowledge regarding biomechanical in vivo deep tissue measures related to spinal manipulation remain somewhat limited. More in vivo animal studies are needed to better understand the effects viscoelastic tissue properties (i.e., dampening) have on applied spinal manipulation forces. This new knowledge may eventually help to determine whether positive clinical outcomes are associated with particular force thresholds reaching superficial and/or deep spinal tissues. A computer-controlled feedback motor and a modified Activator V device with a dynamic load cell attached were used to deliver thrust spinal manipulations at various magnitudes to the L7 spinous process in deeply anesthetized animals. Miniature pressure catheters (Millar SPR-1000) were inserted unilaterally into superficial and deep multifidi muscles. Measurements of applied mechanical forces and superficial/deep multifidi intramuscular pressure changes were recorded during spinal manipulations delivered in vivo. Manipulative forces and net changes in intramuscular pressures reaching deep spinal tissues are greatly diminished by viscoelastic properties of in vivo tissues, which could have possible clinical safety and/or mechanistic implications.

Ex vivo bovine liver nonlinear viscoelastic properties: MR elastography and rheological measurements

IntroductionKnowledge of the nonlinear viscoelastic properties of the liver is important, but the complex tissue behavior outside the linear viscoelastic regime has impeded their characterization, particularly in vivo. Combining static compression with magnetic resonance (MR) elastography has the potential to be a useful imaging method for assessing large deformation mechanical properties of soft tissues in vivo. However, this remains to be verified. Therefore this study aims first to determine whether MR elastography can measure the nonlinear mechanical properties of ex vivo bovine liver tissue under varying levels of uniform and focal preloads (up to 30%), and second to compare MR elastography-derived complex shear modulus with standard rheological measurements. MethodNine fresh bovine livers were collected from a local abattoir, and experiments were conducted within 12hr of death. Two cubic samples (∼10 × 10 × 10 cm3) were dissected from each liver and imaged using MR elastography (60 Hz) under 4 levels of uniform and focal preload (1, 10, 20, and 30% of sample width) to investigate the relationship between MR elastography-derived complex shear modulus (G∗) and the maximum principal Right Cauchy Green Strain (C11). Three tissue samples from each of the same 9 livers underwent oscillatory rheometry under the same 4 preloads (1, 10, 20, and 30% strain). MR elastography-derived complex shear modulus (G∗) from the uniform preload was validated against rheometry by fitting the frequency dependence of G∗ with a power-law and extrapolating rheometry-derived G∗ to 60 Hz. ResultsMR elastography-derived G∗ increased with increasing compressive large deformation strain, and followed a power-law curve (G∗ = 1.73 × C11−0.38, R2 = 0.96). Similarly, rheometry-derived G∗ at 1 Hz, increasing from 0.66 ± 1.03 kPa (1% strain) to 1.84 ± 1.65 kPa (30% strain, RM one-way ANOVA, P < 0.001), and the frequency dependence of G∗ followed a power-law with the exponent decreasing from 0.13 to 0.06 with increasing preload. MR elastography-derived G∗ was 1.4–3.1 times higher than the extrapolated rheometry-derived G∗ at 60 Hz, but the strain dependence was consistent between rheometry and MR elastography measurements. ConclusionsThis study demonstrates that MR elastography can detect changes in ex vivo bovine liver complex shear modulus due to either uniform or focal preload and therefore can be a useful technique to characterize nonlinear viscoelastic properties of soft tissue, provided that strains applied to the tissue can be quantified. Although MR elastography could reliably characterize the strain dependence of the ex vivo bovine liver, MR elastography overestimated the complex shear modulus of the tissue compared to rheological measurements, particularly at lower preload (<10%). That is likely to be important in clinical hepatic MR elastography diagnosis studies if preload is not carefully considered. A limitation is the absence of overlapping frequency between rheometry and MR elastography for formal validation.

Decoupling Uniaxial Tensile Prestress and Waveguide Effects From Estimates of the Complex Shear Modulus in a Cylindrical Structure Using Transverse-Polarized Dynamic Elastography.

Dynamic elastography, whether based on magnetic resonance, ultrasound, or optical modalities, attempts to reconstruct quantitative maps of the viscoelastic properties of biological tissue, properties altered by disease and injury, by noninvasively measuring mechanical wave motion in the tissue. Most reconstruction strategies that have been developed neglect boundary conditions, including quasi-static tensile or compressive loading resulting in a nonzero prestress. Significant prestress is inherent to the functional role of some biological tissues currently being studied using elastography, such as skeletal and cardiac muscle, arterial walls, and the cornea. In the present article a configuration, inspired by muscle elastography but generalizable to other applications, is analytically and experimentally studied. A hyperelastic polymer phantom cylinder is statically elongated in the axial direction while its response to transverse-polarized vibratory excitation is measured. We examine the interplay between uniaxial prestress and waveguide effects in this muscle-like tissue phantom using computational finite element simulations and magnetic resonance elastography measurements. Finite deformations caused by prestress coupled with waveguide effects lead to results that are predicted by a coordinate transformation approach that has been previously used to simplify reconstruction of anisotropic properties using elastography. Here, the approach estimates material viscoelastic properties that are independent of the nonhomogeneous prestress conditions without requiring advanced knowledge of those stress conditions.

Pectin-based bioinks for 3D models of neural tissue produced by a pH-controlled kinetics.

Introduction: In the view of 3D-bioprinting with cell models representative of neural cells, we produced inks to mimic the basic viscoelastic properties of brain tissue. Moving from the concept that rheology provides useful information to predict ink printability, this study improves and expands the potential of the previously published 3D-reactive printing approach by introducing pH as a key parameter to be controlled, together with printing time. Methods: The viscoelastic properties, printability, and microstructure of pectin gels crosslinked with CaCO3 were investigated and their composition was optimized (i.e., by including cell culture medium, HEPES buffer, and collagen). Different cell models representative of the major brain cell populations (i.e., neurons, astrocytes, microglial cells, and oligodendrocytes) were considered. Results and Discussion: The outcomes of this study propose a highly controllable method to optimize the printability of internally crosslinked polysaccharides, without the need for additives or post-printing treatments. By introducing pH as a further parameter to be controlled, it is possible to have multiple (pH-dependent) crosslinking kinetics, without varying hydrogel composition. In addition, the results indicate that not only cells survive and proliferate following 3D-bioprinting, but they can also interact and reorganize hydrogel microstructure. Taken together, the results suggest that pectin-based hydrogels could be successfully applied for neural cell culture.

Biomechanical properties of the stomach: A comprehensive comparative analysis of human and porcine gastric tissue

BackgroundStomach-related disorders impose medical challenges and are associated with significant social and economic costs. The field of biomechanics is promising for understanding tissue behavior and for development of medical treatments and surgical interventions. In gastroenterology, animal models are often used when studies on humans are not possible. Often large animal models with similar anatomical characteristics (size and shape) are preferred. However, it is uncertain if stomachs from humans and large animals have similar mechanical properties. The aim of the present study is to characterize and compare hyper- and viscoelastic properties of porcine and human gastric tissue using tension and radial compression tests. Methods: Hyperelastic and viscoelastic properties were quantified from quasi-static ramp tests and stress relaxation tests. Tension in two directions and radial compression experiments were done on intact stomach wall samples as well as on separated mucosa and muscularis layer samples from porcine and human fundus, corpus and antrum. Results and conclusions: Similar hyper- and viscoelastic constitutive models can be used to describe porcine and human gastric tissue. In total, 19 constitutive parameters were compared and results showed significant variations between species. For example, for intact circumferential samples from antrum, the stiffness (a) and relaxation (τ1) were greater for human samples than for porcine samples (p < 0.0001). The constitutive parameters were condition-, region- and layer-dependent and no distinct pattern hereof between species was found. This indicates that different parameters must be used to describe the specific situation. The present work provides insight into porcine and human gastric radial compressive and tensile hyper- and viscoelastic properties, strengthening the inter-species relation of the biomechanical properties. Constitutive relations were established that may aid development and translation of diagnostic or therapeutic devices with computational models.

Biaxial Tensile Prestress and Waveguide Effects on Estimates of the Complex Shear Modulus Using Optical-Based Dynamic Elastography in Plate-Like Soft Tissue Phantoms.

Dynamic elastography attempts to reconstruct quantitative maps of the viscoelastic properties of biological tissue, properties altered by disease and injury, by noninvasively measuring mechanical wave motion in the tissue. Most reconstruction strategies that have been developed neglect boundary conditions, including quasi-static tensile or compressive loading resulting in a nonzero prestress. Significant prestress is inherent to the functional role of some biological tissues, such as skeletal and cardiac muscle, arterial walls, and the cornea. In the present article a novel configuration, inspired by corneal elastography but generalizable to other applications, is studied. A polymer phantom layer is statically elongated via an in-plane biaxial normal stress while the phantom's response to transverse vibratory excitation is measured. We examine the interplay between biaxial prestress and waveguide effects in this plate-like tissue phantom. Finite static deformations caused by prestressing coupled with waveguide effects lead to results that are predicted by a novel coordinate transformation approach previously used to simplify reconstruction of anisotropic properties. Here, the approach estimates material viscoelastic properties independent of the nonzero prestress conditions without requiring advanced knowledge of those stress conditions.

Proteomic analysis reveals that aging rabbit vocal folds are more vulnerable to changes caused by systemic dehydration

BackgroundOlder adults are more prone to develop systemic dehydration. Systemic dehydration has implications for vocal fold biology by affecting gene and protein expression. The objective of this study was to quantify vocal fold protein changes between two age groups and hydration status, and to investigate the interaction of age and hydration status on protein expression, which has not been investigated in the context of vocal folds before. Comparative proteomics was used to analyze the vocal fold proteome of 6.5-month-old and > 3-year-old rabbits subjected to water ad libitum or water volume restriction protocol.ResultsYoung and older adult rabbits (n = 22) were either euhydrated (water ad libitum) or dehydrated by water volume restriction. Dehydration was confirmed by body weight loss of − 5.4% and − 4.6% in young and older groups, respectively, and a 1.7-fold increase of kidney renin gene expression in the young rabbits. LC-MS/MS identified 2286 proteins in the rabbit vocal folds of young and older adult rabbits combined. Of these, 177, 169, and 81 proteins were significantly (p ≤ 0.05) affected by age, hydration status, or the interaction of both factors, respectively. Analysis of the interaction effect revealed 32 proteins with opposite change patterns after dehydration between older and young rabbit vocal folds, while 31 proteins were differentially regulated only in the older adult rabbits and ten only in the young rabbits in response to systemic dehydration. The magnitude of changes for either up or downregulated proteins was higher in the older rabbits. These proteins are predominantly related to structural components of the extracellular matrix and muscle layer, suggesting a disturbance in the viscoelastic properties of aging vocal fold tissue, especially when subjected to systemic dehydration.ConclusionsWater restriction is a laboratory protocol to assess systemic dehydration-related changes in the vocal fold tissue that is translatable to human subjects. Our findings showed a higher number of proteins differentially regulated with a greater magnitude of change in the vocal folds of older adult rabbits in the presence of systemic dehydration compared to younger rabbits. The association of these proteins with vocal fold structure and biomechanical properties suggests that older human subjects may be more vulnerable to the effects of systemic dehydration on vocal function. The clinical implications of these protein changes warrant more investigation, but age should be taken into consideration when evaluating vocal treatment recommendations that interfere with body fluid balance.

Photoacoustic testing of shear viscoelastic properties of soft tissues using annular beam illumination.

This Letter reports a new, to the best of our knowledge, photoacoustic excitation method for evaluating the shear viscoelastic properties of soft tissues. By illuminating the target surface with an annular pulsed laser beam, circularly converging surface acoustic waves (SAWs) are generated, focused, and detected at the center of the annular beam. The shear elasticity and shear viscosity of the target are extracted from the dispersive phase velocity of the SAWs based on the Kelvin-Voigt model and nonlinear regression fitting. Agar phantoms with different concentrations, and animal liver and fat tissue samples have successfully been characterized. Different from previous methods, the self-focusing of the converging SAWs allows sufficient SNR to be obtained even with low pulsed laser energy density, which makes this approach well compatible with soft tissues under both ex vivo and in vivo testing conditions.

A microfluidic platform to investigate the role of mechanical constraints on tissue reorganization.

Mechanical constraints have a high impact on development processes, and there is a need for new tools to investigate the role of mechanosensitive pathways in tissue reorganization during development. We present here experiments in which embryonic cell aggregates are aspired through constrictions in microfluidic channels, generating highly heterogeneous flows and large cell deformations that can be imaged using two-photon microscopy. This approach provides a way to measure in situ local viscoelastic properties of 3D tissues and connect them to intracellular and intercellular events, such as cell shape changes and cell rearrangements. These methods could be applied to organoids to investigate and quantify rheological properties of tissues, and to understand how constraints affect development.