Torsional wave elastography to assess the mechanical properties of the cornea

Corneal mechanical changes are believed to occur before any visible structural alterations observed during routine clinical evaluation. This study proposed developing an elastography technique based on torsional waves (TWE) adapted to the specificities of the cornea. By measuring the displacements in the propagation plane perpendicular to the axis of the emitter, the effect of guided waves in platelike media was proven negligible. Ex vivo experiments were carried out on porcine corneal samples considering a group of control and one group of alkali burn treatment ( NH 4OH) that modified the mechanical properties. Phase speed was recovered as a function of intraocular pressure (IOP), and a Kelvin-Voigt rheological model was fitted to the dispersion curves to estimate viscoelastic parameters. A comparison with uniaxial tensile testing with thin-walled assumptions was also performed. Both shear elasticity and viscosity correlated positively with IOP, being the elasticity lower and the viscosity higher for the treated group. The viscoelastic parameters ranged from 21.33 to 63.17 kPa, and from 2.82 to 5.30 Pa s, for shear elasticity and viscosity, respectively. As far as the authors know, no other investigations have studied this mechanical plane under low strain ratios, typical of dynamic elastography in corneal tissue. TWE reflected mechanical properties changes after treatment, showing a high potential for clinical diagnosis due to its rapid performance time and paving the way for future in vivo studies.Ministerio de Educacion, Cultura y Deporte Grant DPI2017-83859-R DPI2014-51870-R UNGR15-CE-3664 EQC2018-004508-PSpanish Government DTS15/00093 PI16/00339Instituto de Salud Carlos III y Fondos FederJunta de Andalucia PI-0107-2017 PIN-0030-2017 IE2017-5537MCIN/AEI - European Social Fund "Investing in your future" PRE2018-086085Consejeria de economia, conocimiento, empresas y universidad SOMM17/6109/UGR B-TEP-026- IE2017-5537 P18-RT-1653European Commission SOMM17/6109/UGR B-TEP-026- IE2017-5537 P18-RT-165

Viscoelasticity Imaging of Biological Tissues and Single Cells Using Shear Wave Propagation

Changes in biomechanical properties of biological soft tissues are often associated with physiological dysfunctions. Since biological soft tissues are hydrated, viscoelasticity is likely suitable to represent its solid-like behavior using elasticity and fluid-like behavior using viscosity. Shear wave elastography is a non-invasive imaging technology invented for clinical applications that has shown promise to characterize various tissue viscoelasticity. It is based on measuring and analyzing velocities and attenuations of propagated shear waves. In this review, principles and technical developments of shear wave elastography for viscoelasticity characterization from organ to cellular levels are presented, and different imaging modalities used to track shear wave propagation are described. At a macroscopic scale, techniques for inducing shear waves using an external mechanical vibration, an acoustic radiation pressure or a Lorentz force are reviewed along with imaging approaches proposed to track shear wave propagation, namely ultrasound, magnetic resonance, optical, and photoacoustic means. Then, approaches for theoretical modeling and tracking of shear waves are detailed. Following it, some examples of applications to characterize the viscoelasticity of various organs are given. At a microscopic scale, a novel cellular shear wave elastography method using an external vibration and optical microscopy is illustrated. Finally, current limitations and future directions in shear wave elastography are presented

Applanation optical coherence elastography: noncontact measurement of intraocular pressure, corneal biomechanical properties, and corneal geometry with a single instrument

Current clinical tools provide critical information about ocular health such as intraocular pressure (IOP). However, they lack the ability to quantify tissue material properties, which are potent markers for ocular tissue health and integrity. We describe a single instrument to measure the eye-globe IOP, quantify corneal biomechanical properties, and measure corneal geometry with a technique termed applanation optical coherence elastography (Appl-OCE). An ultrafast OCT system enabled visualization of corneal dynamics during noncontact applanation tonometry and direct measurement of micro air-pulse induced elastic wave propagation. Our preliminary results show that the proposed Appl-OCE system can be used to quantify IOP, corneal biomechanical properties, and corneal geometry, which builds a solid foundation for a unique device that can provide a more complete picture of ocular health

Noncontact elastic wave imaging optical coherence elastography for evaluating changes in corneal elasticity due to crosslinking

The mechanical properties of tissues can provide valuable information about tissue integrity and health and can assist in detecting and monitoring the progression of diseases such as keratoconus. Optical coherence elastography (OCE) is a rapidly emerging technique, which can assess localized mechanical contrast in tissues with micrometer spatial resolution. In this work we present a noncontact method of optical coherence elastography to evaluate the changes in the mechanical properties of the cornea after UV-induced collagen cross-linking. A focused air-pulse induced a low amplitude (μm scale) elastic wave, which then propagated radially and was imaged in three dimensions by a phase-stabilized swept source optical coherence tomography (PhSSSOCT) system. The elastic wave velocity was translated to Young’s modulus in agar phantoms of various concentrations. Additionally, the speed of the elastic wave significantly changed in porcine cornea before and after UV-induced corneal collagen cross-linking (CXL). Moreover, different layers of the cornea, such as the anterior stroma, posterior stroma, and inner region, could be discerned from the phase velocities of the elastic wave. Therefore, because of noncontact excitation and imaging, this method may be useful for in vivo detection of ocular diseases such as keratoconus and evaluation of therapeutic interventions such as CXL

Non-invasive Evaluation of Aortic Stiffness Dependence with Aortic Blood Pressure and Internal Radius by Shear Wave Elastography and Ultrafast Imaging

Elastic properties of arteries have long been recognized as playing a major role in the cardiovascular system. However, non-invasive in vivo assessment of local arterial stiffness remains challenging and imprecise as current techniques rely on indirect estimates such as wall deformation or pulse wave velocity. Recently, Shear Wave Elastography (SWE) has been proposed to non-invasively assess the intrinsic arterial stiffness. In this study, we applied SWE in the abdominal aortas of rats while increasing blood pressure (BP) to investigate the dependence of shear wave speed with invasive arterial pressure and non-invasive arterial diameter measurements. A 15MHz linear array connected to an ultrafast ultrasonic scanner, set non-invasively, on the abdominal aorta of anesthetized rats (N=5) was used. The SWE acquisition followed by an ultrafast (UF) acquisition was repeated at different moment of the cardiac cycle to assess shear wave speed and arterial diameter variations respectively. Invasive arterial BP catheter placed in the carotid, allowed the accurate measurement of pressure responses to increasing does of phenylephrine infused via a venous catheter. The SWE acquisition coupled to the UF acquisition was repeated for different range of pressure. For normal range of BP, the shear wave speed was found to follow the aortic BP variation during a cardiac cycle. A minimum of (5.06$\pm$0.82) m/s during diastole and a maximum of (5.97$\pm$0.90) m/s during systole was measured. After injection of phenylephrine, a strong increase of shear wave speed (13.85$\pm$5.51) m/s was observed for a peak systolic arterial pressure of (190$\pm$10) mmHg. A non-linear relationship between shear wave speed and arterial BP was found. A complete non-invasive method was proposed to characterize the artery with shear wave speed combined with arterial diameter variations. Finally, the results were validated against two parameters the incremental elastic modulus and the pressure elastic modulus derived from BP and arterial diameter variations

Evaluation of posterior porcine sclera elasticity in situ as a function of IOP

The biomechanical properties of the sclera could provide key information regarding the progression and etiology of ocular diseases. For example, an elevated intraocular pressure is one of the most common risk factors for glaucoma and can cause pathological deformations in the tissues of the posterior eye, such as the sclera, potentially damaging these vital tissues. Previous work has evaluated scleral biomechanical response to global displacements with techniques such as inflation testing. However, these methods cannot provide localized biomechanical assessments. In this pilot work, we induce low amplitude (< 10 μm) elastic waves using acoustic radiation force in posterior scleral tissue of fresh porcine eyes (n=2) in situ. The wave propagation induced using an ultrasound transducer was detected across an 8 mm region using a phase-sensitive optical coherence elastography system (PhS-OCE). The elastographic measurements were taken at various artificially controlled intraocular pressures (IOP). The IOP was pre-cycled before being set to 10 mmHg for the first measurement. Subsequent measurements were taken at 20 mmHg and 30 mmHg for each sample. The results show an increase in the stiffness of the sclera as a function of IOP. Furthermore, we observed a variation in the elasticity based on direction, suggesting that the sclera has anisotropic biomechanical properties. Our results show that OCE is an effective method for evaluating the mechanical properties of the sclera, and reveals a new area for our future work

In vivo Optical Coherence Elastography Reveals Spatial Variation and Anisotropy of Corneal Stiffness

Objective: The mechanical properties of corneal tissues play a crucial role in determining corneal shape and have significant implications in vision care. This study aimed to address the challenge of obtaining accurate in vivo data for the human cornea. Methods: We have developed a high-frequency optical coherence elastography (OCE) technique using shear-like antisymmetric (A0)-mode Lamb waves at frequencies above 10 kHz. Results: By incorporating an anisotropic, nonlinear constitutive model and utilizing the acoustoelastic theory, we gained quantitative insights into the influence of corneal tension on wave speeds and elastic moduli. Our study revealed significant spatial variations in the shear modulus of the corneal stroma on healthy subjects for the first time. The central cornea exhibited a shear modulus of 74 kPa, while the corneal periphery showed a decrease to 41 kPa. The limbus demonstrated an increased shear modulus exceeding 100 kPa. We obtained wave displacement profiles that are consistent with highly anisotropic corneal tissues. Conclusion: Our approach enabled precise measurement of corneal tissue elastic moduli in situ with high precision (< 7%) and high spatial resolution (< 1 mm). Significance: The high-frequency OCE technique holds promise for biomechanical evaluation in clinical settings, providing valuable information for refractive surgeries, degenerative disorder diagnoses, and intraocular pressure assessments

Possible depth-resolved reconstruction of shear moduli in the cornea following collagen crosslinking (CXL) with optical coherence tomography and elastography

Corneal collagen crosslinking (CXL) is commonly used to prevent or treat keratoconus. Although changes in corneal stiffness induced by CXL surgery can be monitored with non-contact dynamic optical coherence elastography (OCE) by tracking mechanical wave propagation, depth dependent changes are still unclear if the cornea is not crosslinked through the whole depth. Here, phase-decorrelation measurements on optical coherence tomography (OCT) structural images are combined with acoustic micro-tapping (A$\mu$T) OCE to explore possible reconstruction of depth-dependent stiffness within crosslinked corneas in an ex vivo human cornea sample. Experimental OCT images are analyzed to define the penetration depth of CXL into the cornea. In a representative ex vivo human cornea sample, crosslinking depth varied from $\sim 100\mu m$ in the periphery to $\sim 150\mu m$ in the cornea center and exhibited a sharp in-depth transition between crosslinked and untreated areas. This information was used in an analytical two-layer guided wave propagation model to quantify the stiffness of the treated layer. We also discuss how the elastic moduli of partially CXL-treated cornea layers reflect the effective engineering stiffness of the entire cornea to properly quantify corneal deformation.Comment: Submitted to Biomedical Optics Express on June 13th 2023, Manuscript ID: 497970 - Under Review. Manuscript, 10 pages / 6 figures / 2 tables. Supplementary, 7 pages / 4 figure

Possible depth-resolved reconstruction of shear moduli in the cornea following collagen crosslinking (CXL) with optical coherence tomography and elastography

Collagen crosslinking of the cornea (CXL) is commonly employed to prevent or treat keratoconus. Although the change of corneal stiffness induced by CXL surgery can be monitored with non-contact dynamic Optical Coherence Elastography (OCE) by tracking mechanical wave propagation, the depth dependence of this change is still unclear if the cornea is not crosslinked through the whole depth. Here we propose to combine phase-decorrelation measurement applied to OCT structural images and acoustic micro-tapping (A$\mu$T) OCE to explore possible depth reconstruction of stiffness within crosslinked corneas in an ex vivo human cornea sample. The analysis of experimental OCT images is used to define the penetration depth of CXL into the cornea, which varies from $\sim$100$\mu m$ in the periphery to $\sim$150$\mu m$ in the central area and exhibits a sharp transition between areas. This information was used in a two-layer analytical model to quantify the stiffness of the treated layer. We also discuss how the elastic moduli of partially CXL-treated cornea layers reconstructed from OCE measurements reflect the effective mechanical stiffness of the entire cornea to properly quantify surgical outcome.Comment: Main: 10 Pages, 6 Figures Supplemental: 12 Pages, 3 Figure

Development and Applications of Advanced Ultrasound Techniques for Characterization and Stimulation of Engineered Tissues

Mechanobiology is central in the development, pathology, and regeneration of musculoskeletal tissues, in which mechanical factors play important roles. Therefore, there is a need for methods to characterize the composition and mechanical properties of developing musculoskeletal tissues over time. Ultrasound elastographic techniques have been developed for noninvasive imaging of spatial heterogeneity in tissue stiffness. However, their application for quantitative assessment of tissue mechanical properties, especially viscoelastic properties, has not been exploited. Additionally, ultrasound energy may be used to apply mechanical stimulation to engineered constructs at the microscale, and thereby to enhance tissue regeneration. We have developed a multimode ultrasound viscoelastography (MUVE) system for assessing microscale mechanical properties of engineered hydrogels. MUVE uses focused ultrasound pulses to apply acoustic radiation force (ARF) to deform samples, while concurrently measuring sample dimensions using coaxial high frequency ultrasound imaging. We used MUVE to perform creep tests on agarose, collagen, and fibrin hydrogels of defined concentrations, as well as to monitor the mechanical properties of cell-seeded constructs over time. Local and bulk viscoelastic properties were extracted from strain-time curves through fitting of relevant constitutive models, showing clear differences between concentrations and materials. In particular, we showed that MUVE is capable of mapping heterogeneity of samples in 3D. Using inclusion of dense agarose microbeads within agarose, collagen and fibrin hydrogels, we determined the spatial resolution of MUVE to be approximately 200 μm in both the lateral and axial directions. Comparison of MUVE to nanoindentation and shear rheometry showed that our ultrasound-based technique was superior in generating consistent, microscale data, particularly for very soft materials. We have also adapted MUVE to generate localized cyclic compression, as a means to mechanically stimulate engineered tissue constructs at the microscale. Selected treatment protocols were shown to enhance the osteogenic differentiation of human mesenchymal stem cells in collagen-fibrin hydrogels. Constructs treated at 1 Hz at an acoustic pressure of 0.7 MPa for 30 minutes per day showed accelerated osteogenesis and increased mineralization by 10 to 30 percent, relative to unstimulated controls. In separate experiments, the ultrasound pulse intensity was increased over time to compensate for changes in matrix properties over time, and a 35 percent increase in mineralization was achieved. We also extended the application of a previously-developed spectral ultrasound imaging (SUSI) technique to an animal model for early detection of heterotopic ossification (HO). The quantitative information on acoustic scatterer size and concentration derived from SUSI was used to differentiate tissue composition in a burn/tenotomy mice model from the control model. Importantly, HO foci were detected as early as one week after injury using SUSI, which is 3-5 weeks earlier than when using conventional micro-computed tomography. Taken together, these results demonstrate that ultrasound-based techniques can non-invasively and quantitatively characterize viscoelastic properties of soft materials in 3D, as well as their composition over time. Ultrasound pulses can also be used to stimulate engineered constructs to promote musculoskeletal tissue formation. MUVE, SUSI, and ultrasound stimulation can be combined into an integrated system to investigate the roles of matrix composition, static mechanical environment, and dynamic mechanical stimuli in tissue regeneration, as well as the interactions of these factors and their evolution over time. Ultrasound-based techniques therefore have promising potential in noninvasively characterizing the composition and biomechanics, as well as providing mechanical intervention in native and engineered tissues as they develop over time.PHDBiomedical EngineeringUniversity of Michigan, Horace H. Rackham School of Graduate Studieshttps://deepblue.lib.umich.edu/bitstream/2027.42/144116/1/xho_1.pd