SignificanceThe skin’s mechanical properties are tightly regulated. Various pathologies can affect skin stiffness, and understanding these changes is a focus in tissue engineering. Ex vivo skin scaffolds are a robust platform for evaluating the effects of various genetic and molecular interactions on the skin. Transforming growth factor-beta (TGF-β) is a critical signaling molecule in the skin that can regulate the amount of collagen and elastin in the skin and, consequently, its mechanical properties.AimThis study investigates the biomechanical properties of bio-engineered skin scaffolds, focusing on the influence of TGF-β, a signaling molecule with diverse cellular functions.ApproachThe TGF-β receptor I inhibitor, galunisertib, was employed to assess the mechanical changes resulting from dysregulation of TGF-β. Skin scaffold samples, grouped into three categories (control, TGF-β-treated, and TGF-β + galunisertib-treated), were prepared in two distinct culture media—one with aprotinin (AP) and another without. Two optical elastography techniques, namely wave-based optical coherence elastography (OCE) and Brillouin microscopy, were utilized to quantify the biomechanical properties of the tissues.ResultsResults showed significantly higher wave speed (with AP, p<0.001; without AP, p<0.001) and Brillouin frequency shift (with AP, p<0.001; without AP, p=0.01) in TGF-β-treated group compared with the control group. The difference in wave speed between the control and TGF-β + galunisertib with (p=0.10) and without AP (p=0.36) was not significant. Moreover, the TGF-β + galunisertib-treated group exhibited lower wave speed without and with AP and reduced Brillouin frequency shift than the TGF-β-treated group without AP, further strengthening the potential role of TGF-β in regulating the mechanical properties of the samples.ConclusionsThese findings offer valuable insights into TGF-β-induced biomechanical alterations in bio-engineered skin scaffolds, highlighting the potential of OCE and Brillouin microscopy in the development of targeted therapies in conditions involving abnormal tissue remodeling and fibrosis.
Assessing the biomechanical properties of tissues can have important applications for disease diagnosis and treatment monitoring. Optical coherence elastography is an established technology for measuring the biomechanical properties of tissues, and has been implemented ex vivo, in vivo, and clinically. Various steps are required for effective translation of this technology, including making improvements in data analysis and processing. OCE data can be inherently noisy, a single data acquisition can consist of gigabytes of data, and data processing can be lengthy. In this work, we examine how convolutional neural networks can be implemented to efficiently process OCE data.
The biomechanical properties of the cornea are tightly linked to its structure. Refractive procedures, including laser assisted in situ keratomileusis (LASIK), can be used to correct myopia and hyperopia through precision alteration of corneal structure. However, structural alterations undoubtedly have consequences for its biomechanical integrity. Here, we utilize optical coherence elastography (OCE) to evaluate changes in corneal biomechanical properties after LASIK in ex vivo porcine corneas using three techniques, air coupled ultrasound OCE (ACUS-OCE), heartbeat OCE (Hb-OCE), and compression OCE. The results suggest that LASIK reduces tissue stiffness. Future work will examine stiffness changes in vivo.
Assessing corneal biomechanical properties may be important for the diagnosis of ocular diseases as the mechanical properties of the cornea change during disease development. Elastography is a technique to image the mechanical properties of tissues by applying a mechanical load to the tissue and measuring the resultant displacement using available imaging techniques like magnetic resonance or ultrasound imaging. The measured displacement is then translated to mechanical properties. Heartbeat optical coherence elastography (Hb-OCE) is a completely passive elastography technique, which uses physiological perturbations naturally present in the body in lieu of active tissue stimulation sources. In this work, we demonstrate the first use of 3D Hb-OCE to measure the biomechanical properties of the cornea in 3D in an ex vivo porcine eye. Measurements were taken on whole porcine eye globes using a Fourier Domain Mode Locked swept source laser-based OCT system with a volume rate of 4.2 Hz. The strain in the cornea was measured between successive B-scans during a simulated ocular pulse, and each scan was stacked together to obtain 3D volumetric strain due to the simulated heartbeat. This technique may potentially enable full volumetric analysis of corneal mechanical properties completely passively. Future work will focus on 3D evaluation of customized corneal crosslinking and in vivo translation of the 3D Hb-OCE technique.
Assessing corneal biomechanical properties is useful for the diagnosis of ocular disease and for monitoring therapeutic intervention. Here, we demonstrate how two different forms of elastography can be combined to perform a multimodal biomechanical analysis of the cornea. Heartbeat optical coherence elastography (Hb-OCE) and compression elastography were combined to measure the mechanical properties of rabbit corneas in vivo. The results demonstrate that the difference in corneal stiffness measured by Hb-OCE and compression OCE is statistically insignificant, suggesting that both techniques could potentially be used interchangeably to measure corneal stiffness.
Measuring the mechanical properties of the cornea can help understand the structure and physiology of the eye, early detection of disease, and evaluation of therapy outcomes. In this work, we investigate the effect of collagen XII deficiency on the stiffness of the murine cornea using a multimodal approach for biomechanical analysis. Wave-based optical coherence elastography (OCE), heartbeat OCE, and Brillouin microscopy were all utilized to assess the mechanical properties of wild-type and collagen XII deficient ex vivo murine corneas as a function of IOP. All three techniques show that collagen XII deficiency leads to a dramatic decrease in corneal stiffness. Future work will investigate how these measurement techniques can be translated for in vivo assessment of corneal elasticity to understand the contribution of various proteins to corneal structural and mechanical integrity.
Measuring corneal biomechanical properties provides important diagnostic information regarding tissue health and for evaluating the outcomes of therapies. Heartbeat OCE (Hb-OCE), a truly passive elastography technique, utilizes the pulsatile nature of the intraocular pressure (IOP) to quantify corneal stiffness completely noninvasively and with no external excitation. In this work, we utilized whole rabbit eye globes to quantify the displacement and strain in the cornea by fluctuating the IOP ex-vivo. We also approximated the non-linearity of the stiffness of the cornea by performing HbOCE measurements at various baseline IOPs. The results show an expected increase in corneal stiffness as the baseline IOP increased, demonstrating the effectiveness of Hb-OCE as a tool for measuring corneal biomechanical properties completely noninvasively and with no external excitation.
We present an ultra-fast single-shot line-field optical coherence elastography (LFOCE) technique based on a parallel line-field spectral domain optical coherence tomography system. The system was based on a Michelson-type interferometer, a supercontinuum broadband light source, and an ultra-fast 2D spectrometer. The peak sensitivity was ~102 dB, and the sensitivity roll-off was ~37 dB over 0.8 mm. The 1/e length along the line beam as measured by the signal to noise ratio (SNR) from an image of a mirror was 3.1 mm. The 1/e width across the line beam was measured by the knife-edge technique and was ~9.7 μm. The displacement stability as measured by standard deviation over 20 ms of a glass coverslip in common-path mode was 0.52 nm at an OCT SNR of 41 dB. The camera operated at a framerate of 25 kHz with 460 lateral pixels, resulting in an A-scan rate of 11.5 MHz. Validation was performed in gelatin phantoms of various concentrations, and the results corroborated well with mechanical testing. After validation in the phantoms, OCE measurements were performed in rabbit corneas in situ in the whole eye-globe configuration. The eyes were cannulated for artificial intraocular pressure (IOP) control. The elastic wave speeds in the cornea at 10, 15, and 20 mmHg were 3.03±0.05, 4.66±0.03, and 8.85±0.08 m/s, respectively. OCE measurements were also performed in an in vivo anesthetized rabbit, and the wave propagation was successfully captured. These results show the ability of the ultra-fast OCE system to measure changes in stiffness as well performing live measurements.
Currently available clinical devices cannot provide localized maps of corneal stiffness. We present a phase-sensitive method of compressional optical coherence elastography (OCE) for mapping corneal stiffness. In situ measurements showed that the strain, ε, slightly decreased as a function of IOP and CXL significantly decreased the strain (P<0.001). The change in pressure in the eye-globe, ΔIOP, was also measured during compression. We quantified the elasticity, E, of the cornea by E=ΔIOP/ε. In contrast to the strain, there was a significant effect of the baseline IOP on the stiffness (P<0.001). After CXL, there was a ~85% increase in stiffness, which was significant (P<0.001). In addition to the whole sample CXL, partial CXL was performed where only one half of the cornea was treated. The strain in the untreated region was significantly greater (P=0.030) than the strain in the CXL region, and the elasticity was significantly greater in the CXL region (P=0.030). Next, in vivo measurements were performed in an anesthetized rabbit. The strain decreased by ~75% after CXL, which was significant (P<0.001). Partial CXL was performed on another animal, where only half the cornea was treated. The strain in the untreated region was significantly less (P<0.001) than the strain in the CXL region. Our results show the capability of compression-based OCE to measure changes in corneal biomechanical properties in 4 different scenarios (in situ traditional CXL, in situ partial CXL, in vivo traditional CXL, in vivo partial CXL).
Corneal stiffness is an important parameter in assessing tissue health. Here, we demonstrate Hb-OCE to measure the stiffness of the rabbit cornea in vivo as a function of the ocular pulse.
The purpose of this study is to evaluate the changes in elasticity of the corneo-scleral limbus when the eye-globe is subjected to different intraocular pressures. Continuous elastic waves were mechanically induced in the sclera and imaged using a phase-sensitive optical coherence tomography system. Experimental results showed a marked heterogeneity of wave speed across the corneal-limbal-scleral region, which increased as a function of IOP. The conclusion of this study reveals that the corneo-scleral limbus tissue is more mechanically nonlinear compared to the cornea and sclera. This suggests that limbus acts as a supporting and accommodating mechanical structure for the cornea.
Corneal biomechanical properties are useful for the diagnosis of ocular diseases and monitoring of therapeutic interventions. Elastography is one method of assessing corneal biomechanical properties but typically relies on an active source of displacement. Here, we present a passive elastography technique known as heartbeat optical coherence elastography (Hb-OCE), where corneal stiffness is quantified by measuring its response to the ocular pulse. The technique was performed before and after crosslinking, which stiffens the cornea. First, Hb-OCE was performed in situ to assess the feasibility of the technique. Next, the technique was performed in vivo, suggesting that Hb-OCE may be feasible for clinical assessment of corneal stiffness without the need for any contact or external excitation.
Corneal biomechanical properties can provide important information to measure tissue health. Elastography provides a useful tool to measure the biomechanical properties of tissues in vivo, but typically relies on measuring a response to external excitation. Passive elastography is a method to assess mechanical response to physiological pressures, which can be useful in the field of ophthalmology. In this work, we utilize our technique, Hb-OCE, to measure the mechanical properties of cornea in vivo. Furthermore, we were able to successfully distinguish stiffness in an untreated and crosslinked cornea using this technique, demonstrating its effectiveness as a tool for measuring corneal biomechanics.
We have designed and developed an air-coupled ultrasonic radiation force probe co-focused with a phase-sensitive optical coherence tomography system (ACUS-OCT). Our custom-made 1 MHz spherically focused piezo-electric transducer with a concentric circular opening of 10 mm diameter allows for the confocal micro-excitation of waves and the spatial (2D, 3D) motion measurement of tissues. Phantom studies demonstrated the capabilities of this probe to produce quasi-harmonic excitation up to 4 kHz for the generation of highly localized elastic waves. Experimental results in ocular tissues showed the highly localized 2D/3D elasticity mapping capabilities of this approach with great potential for clinical translation.
Significance: Mechanical assessment of the cornea can provide important structural and functional information regarding its health. Current clinically available tools are limited in their efficacy at measuring corneal mechanical properties. Elastography allows for the direct estimation of mechanical properties of tissues in vivo but is generally performed using external excitation force.
Aim: To show that heartbeat optical coherence elastography (Hb-OCE) can be used to assess the mechanical properties of the cornea in vivo.
Approach: Hb-OCE was utilized to detect Hb-induced deformations in the rabbit cornea in vivo without the need for external excitation. Furthermore, we demonstrate how this technique can distinguish corneal stiffness between untreated (UT) and crosslinked (CXL) tissue.
Results: Our results demonstrate that stiffness changes in the cornea can be detected using only the Hb-induced deformations in the cornea. Additionally, we demonstrate a statistically significant difference in strain between the UT and CXL corneas.
Conclusions: Hb-OCE may be an effective tool for assessing the mechanical properties of the cornea in vivo without the need for external excitation. This tool may be effective for clinical assessment of corneal mechanical properties because it only requires optical coherence tomography imaging and data processing.
Significance: It is generally agreed that the corneal mechanical properties are strongly linked to many eye diseases and could be used to assess disease progression and response to therapies. Elastography is the most notable method of assessing corneal mechanical properties, but it generally requires some type of external excitation to induce a measurable displacement in the tissue.
Aim: We present Heartbeat Optical Coherence Elastography (Hb-OCE), a truly passive method that can measure the elasticity of the cornea based on intrinsic corneal displacements induced by the heartbeat.
Approach: Hb-OCE measurements were performed in untreated and UV-A/riboflavin cross-linked porcine corneas ex vivo, and a distinct difference in strain was detected. Furthermore, a partially cross-linked cornea was also assessed, and the treated and untreated areas were similarly distinguished.
Results: Our results suggest that Hb-OCE can spatially map displacements in the cornea induced by small fluctuations in intraocular pressure, similar to what is induced by the heartbeat.
Conclusions: The described technique opens the possibility for completely passive and noncontact in vivo assessment of corneal stiffness.
Quantifying the biomechanical properties of the cornea is required for accurate assessment of tissue health. In this work, we evaluate cornea biomechanical properties during intraocular pressure fluctuation. This method results in a more physiologically relevant assessment of tissue mechanics, and thus diverges from typical elastography techniques that use externally induced deformation. Optical coherence elastography is used to detect displacements in the cornea as IOP is cycled during the heartbeat. The axial strains within the cornea are calculated from these displacements, so that dynamic changes in stiffness can be determined. The results indicate a gradient in axial strain within the cornea.
Age-related cataract is one of the most prevalent causes of visual impairment worldwide. Early detection of cataracts can be immensely helpful for preserving visual acuity by ensuring that the appropriate therapeutic procedures are performed at earlier stages of the disease. In this work, we investigated the relationship between the progression of oxidative cataract and the biomechanical properties of the crystalline lens. We assessed the changes in the stiffness induced by cataracts in porcine lenses in vitro with dynamic optical coherence elastography. The efficacy of α-lipoic acid to minimize the stiffening of the lens was also quantified. The results showed a significant increase in Young’s modulus of the lens due to the formation of the oxidative cataract (from ~ 8 kPa to ~123 kPa). Young’s moduli of the lenses decreased after incubation in α-Lipoic Acid (~123 kPa vs ~45 kPa). These results show that the lens stiffness increased during oxidative cataract formation and that α-lipoic acid has the potential to reduce the stiffening of the lens caused by the oxidative damage.
The biomechanical properties of the crystalline lens play a crucial role in its visual function. Assessing biomechanical properties of the lens may help with early disease detection and robust assessment of therapeutic interventions. However, measuring the biomechanical properties of the lens is a challenge due to its location inside the eye-globe. In this study, we demonstrate the combination of optical coherence elastography (OCE) and Brillouin microscopy to evaluate the stiffness of porcine lenses ex vivo (N=6). Brillouin microscopy can map the Brillouin-derived longitudinal modulus of the whole lens, but imaging times are lengthy. OCE can provide quantitative measurements of viscoelasticity rapidly, but the limited scattering of the lens limits its in-depth measurements. By combining these two techniques, we show a strong correlation between the Brillouin modulus and OCE-measured Young’s modulus in the lens, enabling depth-wise mapping of the Young’s modulus. The correlation coefficient between the two measurements was R=0.89. Using this correlation, the elasticity of the anterior lens was 2.72±0.89 kPa, and the mean Young’s modulus of the nucleus was 12.92±2.75 kPa. Similarly, the elasticity of the posterior lens was 3.80±1.25 kPa. While both techniques can evaluate the stiffness of the biological tissues separately, our work demonstrates that combining these techniques could enable mapping of the Young’s modulus completely noninvasively in non-scattering tissues such as the crystalline lens.
The biomechanical properties of the cornea can be an important biomarker for assessing tissue health. In this work, we evaluated cornea biomechanical properties during intraocular pressure fluctuation simulating heartbeat induced corneal pulse. This method results in a more physiologically relevant assessment of tissue mechanics, and thus diverges from typical elastography techniques that rely on externally induced deformation. Optical coherence tomography is used to detect displacements in the cornea as IOP is cycled. The axial strains within the cornea are calculated from these displacements, so that dynamic changes in stiffness can be determined. The results indicate a gradient in axial displacement within the cornea and illustrate a distinct difference in strain between the untreated and crosslinked tissues. This suggests that heartbeat OCE may be a useful technique in assessing stiffness of the cornea.
Cataract is one of the most prevalent causes of blindness around the world. Understanding the mechanisms of cataract development and progression is important for clinical diagnosis and treatment. Cold cataract has proven to be a robust model for cataract formation that can be easily controlled in the laboratory. There is evidence that the biomechanical properties of the lens can be significantly changed by cataract. Therefore, early detection of cataract, as well as evaluation of therapies, could be guided by characterization of lenticular biomechanical properties. In this work, we utilized optical coherence elastography (OCE) to monitor the changes in biomechanical properties of ex vivo porcine lenses during formation of cold cataract. Elastic waves were induced in the porcine lenses by a focused micro air-pulse while the lenses were cooled, and the elastic wave velocity was translated to Young’s modulus of the lens. The results show an increase in the stiffness of the lens due to formation of the cold cataract (from 11.3 ± 3.4 to 21.8 ± 7.8 kPa). These results show a relation between lens opacity and stiffness and demonstrate that OCE can assess lenticular biomechanical properties and may be useful for detecting and potentially characterizing cataracts.
Wave-based optical coherence elastography (OCE) is a rapidly emerging technique for localized elasticity assessment of tissues due to its high displacement sensitivity and simple implementation. This method does not require prior knowledge of mechanical load characteristics, such as the applied preload and applied stress on the sample. Currently, noncontact wave excitation has been accomplished with various methods, such as focused micro air-pulse and acoustic techniques. However, they are limited by the inability to target specific tissues and usually only image the transversely propagating elastic wave, which generally requires scanning the probe beam across the sample. In addition, the upper frequency components of the elastic waves are limited to a few kilohertz, which are sensitive to boundary conditions due to their long wavelengths. In this study, we demonstrated that rapid vaporization of perfluorocarbon inside dye nanoparticles that was excited by a pulsed laser excitation, termed “nanobombs”, can produce high frequency longitudinal elastic waves in tissue mimicking phantoms. The nanoparticles were excited by a 1064 nm pulsed laser, which was co-focused with the OCT probe beam. The longitudinal elastic waves, which propagated axially (i.e., following the optical path), were directly imaged by a phase-sensitive Fourier domain mode-locked based OCT system. The detected elasticity was validated with well-established air-pulse OCE and the “gold standard” uniaxial mechanical testing. The results demonstrate the feasibility of performing nanobomb elastography in tissue with the potential for targeting specific tissues and producing longitudinal elastic waves with high frequency content.
Colon pathologies including colon cancer and ulcerative colitis afflict hundreds of thousands of people in the United States. Clinical detection of colon diseases is generally performed through colonoscopy. However, these methods usually lack the sensitivity or resolution to detect diseased tissue at early stages. Even high resolution optical techniques such as confocal microscopy and optical coherence tomography (OCT) rely on structural features to detect anomalies in tissue, which may not be sufficient for early disease detection. If changes in tissue biomechanical properties precede morphological changes in tissue physiology, then mechanical contrast would enable earlier detection of disease. In this work, we utilized optical coherence elastography (OCE) to assess the biomechanical properties of healthy, cancerous, and colitis tissue. Additionally, the optical properties of each sample were also assessed as a secondary feature to distinguish tissue types. The Young’s modulus, as measured by the propagation of an elastic wave, of the healthy, cancerous, and colitis tissue was 10.8 ± 1.0 kPa, 7.12 ± 1.0 kPa, and 5.1 ± 0.1 kPa, respectively. The variations in the OCT signal intensity over depth, as measured by the slope-removed standard deviation of each A-scan was 5.8 ±.0.3 dB, 5.1 ± 0.4 dB, and 5.5 ± 0.2 dB for healthy, cancerous, and colitis tissue, respectively. This work shows OCT structural imaging combined with OCE can detect minute changes in colon tissue optical scattering and elastic properties, which may be useful for detection various colon diseases, such as colitis and colon cancer.
Wave-based optical elastography is a rapidly emerging technique for viscoelastic assessment of tissues due to its high displacement sensitivity and simple implementation. This method does not require prior knowledge of mechanical load characteristics, such as the applied preload and applied stress on the sample. However, current truly noncontact excitation methods are limited by their inability to produce broadband waves with high frequency content. Lower frequency wave content is constrained by boundary conditions, and thus, requires specifically tailored mechanical models that consider the sample geometry. In this work, we demonstrate that rapid vaporization of perfluorocarbon inside dye nanoparticles (NP) with a pulsed laser can produce high frequency and broadband elastic waves in tissue mimicking agar phantoms. As a comparison, a focused air-pulse was used as an alternative excitation method. The elastic waves were imaged by an ultra-fast low-coherence line-field holography system. Our results show that the NPs produced elastic waves with frequencies up to ~9 kHz, while the air-pulse was only able to produce waves with frequency content up to ~2 kHz. The elastic wave dispersion curves were fitted to the analytical solution of a Rayleigh wave model to quantify viscoelasticity. Analysis of the broadband high-frequency waves produced by the NPs yielded more accurate quantification of the sample viscoelasticity, demonstrating the benefits of optically excited elastic waves.
Screening for ocular diseases, such as glaucoma and keratoconus, includes measuring the eye-globe intraocular pressure (IOP) and corneal biomechanical properties. However, currently available clinical tools cannot quantify corneal tissue material parameters, which can provide critical information for detecting diseases and evaluating therapeutic outcomes. Here, we demonstrate measurement of eye-globe IOP, corneal elasticity, and corneal geometry of in situ porcine corneas with a technique termed applanation optical coherence elastography (Appl-OCE) with single instrument. We utilize an ultrafast phase-sensitive optical coherence tomography system comprised of a 4X buffered Fourier domain mode-locked swept source laser with an Ascan rate of ~1.5 MHz and a 7.3 kHz resonant scanner. The IOP was measured by imaging the response of in situ porcine corneas to a large force air-puff. As with other noncontact tonometers, the time when the cornea was applanated during the inwards and outwards motion was correlated to a measure air-pressure temporal profile. The IOP was also measured with a commercially available rebound tonometer for comparison. The stiffness of the corneas was assessed by directly imaging and analyzing the propagation of a focused micro air-pulse induced elastic wave, and the corneal geometry was obtained from the OCT structural image. Our results show that corneal thickness decreased as IOP increased, and that corneal stiffness increased with IOP. Moreover, the IOP measurements made by Appl-OCE were more closely correlated with the artificially set IOP than the rebound tonometer, demonstrating the capabilities of Appl-OCE to measure corneal stiffness, eye-globe IOP, and corneal geometry with a single instrument.
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.
The mechanophysiology of tissues in the posterior eye have been implicated for diseases such as myopia and glaucoma. For example, the eye-globe shape, and consequently optical axial length, can be affected by scleral stiffness. In glaucoma, an elevated intraocular pressure is the primary risk factor for glaucoma, which is the 2nd most prevalent known cause of blindness. Recent work has shown that biomechanical properties of the optic nerve are critical for the onset and progression of glaucoma because weak tissues cause large displacements in the optic nerve, causing tissue damage. In this work, we utilize air-pulse optical coherence elastography (OCE) to quantify the spatial distribution of biomechanical properties of the optic nerve, its surrounding tissues, and the posterior sclera. Air-pulse measurements were made in a grid on in situ porcine eyes in the whole eye-globe configuration as various IOPs. The OCE-measured displacement process was linked to tissue stiffness by a simple kinematic equation. The results show that the optic nerve and peripapillary sclera are much stiffer than the surrounding sclera, and the stiffness of the optic nerve and peripapillary sclera increased as a function of IOP. However, the stiffness of the surrounding sclera did not dramatically increase. Our results show that understanding the dynamics of the biomechanical properties of the eye are critical to understand the aforementioned diseases and may provide additional information for assessing visual health and integrity.
The collagen fibril orientation of the cornea can provide critical information about cornea tissue health because diseases such as keratoconus and therapeutic interventions such as UV-A/riboflavin corneal collagen crosslinking (CXL) can alter the ultrastructural arrangement of collagen fibrils. Here, we quantify the elastic anisotropy and hysteresis of in situ porcine corneas as a function of intraocular pressure (IOP) with noncontact optical coherence elastography. Moreover, the effects of UV-A riboflavin corneal collagen crosslinking on the elastic anisotropy and hysteresis were evaluated. The propagation of an air-pulse induced elastic wave was imaged at stepped meridional angles by a home built phasestabilized swept source OCE system. The stiffness of the cornea was translated from the velocity of the wave, and the elastic anisotropy was quantified by modifying the planar anisotropy coefficient. As the IOP increased, the stiffness of the corneas increased from ~18 kPa at 15 mmHg IOP to ~ 120 kPa at 30 mmHg IOP. While there was a measureable hysteresis, it was not significant. After CXL, the Young’s modulus of the corneas significantly increased from ~18 kPa to ~44 kPa at 15 mmHg IOP. The mechanical anisotropy also increased significantly from ~10 a.u. in the untreated corneas to ~23 a.u. in the CXL treated corneas, 15 mmHg IOP. However, CXL did not change the elastic anisotropic orientation, and the mechanical anisotropic hysteresis was not significant after CXL.
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.
Recent work has shown that the biomechanical properties of tissues in the posterior eye have are critical for
understanding the etiology and progression of ocular diseases. For instance, the primary risk for glaucoma is an elevated
intraocular pressure (IOP). Weak tissues will deform under the large pressure, causing damage to vital tissues. In
addition, scleral elasticity can influence the shape of the eye-globe, altering the axial length. In this work, we utilize a
noncontact form of optical coherence elastography (OCE) to quantify the spatial distribution of biomechanical properties
of the optic nerve, its surrounding tissues, and posterior sclera on the exterior of in situ porcine eyes in the whole eyeglobe
configuration. The OCE measurements were taken at various IOPs to evaluate the biomechanical properties of the
tissues as a function of IOP. The air-pulse induced dynamic response of the tissues was linked to Young’s modulus by a
simple kinematic equation by quantified the damped natural frequency (DNF). The results show that the posterior sclera
is not as stiff as the optic nerve and its surrounding tissues (~460 Hz and ~894 Hz at 10 mmHg IOP, respectively).
Moreover, the scleral stiffness was generally unaffected by IOP (~460 Hz at 10 mmHg IOP as compared to ~516 Hz at
20 mmHg), whereas the optic nerve and its surrounding tissues stiffened as IOP was increased (~894 Hz at 10 mmHg to
~1221 Hz at 20 mmHg).
Diseases such as keratoconus can alter the orientation of collagen fibrils in the cornea. Moreover, therapeutic
interventions such as UV-A/riboflavin corneal collagen crosslinking (CXL) can alter the collagen fibril arrangement.
Therefore, the anisotropic characteristics of the cornea can provide vital information about tissue integrity. In this work,
we utilize noncontact elastic wave imaging optical coherence elastography (EWI-OCE) to assess the elastic anisotropy
and hysteresis of in situ porcine corneas as various intraocular pressures (IOP). In addition, we evaluated the effects of
CXL on the mechanical anisotropy and hysteresis. OCE measurements were made at stepped meridional angles, and a
sliding window algorithm spatially mapped the elasticity. A modified planar anisotropy coefficient was utilized to
quantify the elastic anisotropy of the corneas. The results show that the stiffness and elastic anisotropy of the corneas
were significantly affected by CXL and IOP (P<0.001), but the hysteresis was not significant (P<0.05). Moreover, the
changes in elasticity due to CXL were angle-dependent (P<0.005). However, the changes in mechanical anisotropy from
CXL were not angle-dependent (P>0.05).
This study demonstrates the feasibility of using the Rayleigh wave model (RWM) in combination with optical coherence elastography (OCE) technique to assess the viscoelasticity of soft tissues. Dispersion curves calculated from the spectral decomposition of OCE-measured air-pulse induced elastic waves were used to quantify the viscoelasticity of samples using the RWM. Validation studies were first conducted on 10% gelatin phantoms with different concentrations of oil. The results showed that the oil increased the viscosity of the gelatin phantom samples. This method was then used to quantify the viscoelasticity of chicken liver. The Young’s modulus of the chicken liver tissues was estimated as E=2.04±0.88 kPa with a shear viscosity η=1.20±0.13 Pa s. The analytical solution of the RWM correlated very well with the OCE-measured phased velocities (R2=0.96±0.04). The results show that the combination of the RWM and OCE is a promising method for noninvasively quantifying the biomechanical properties of soft tissues and may be a useful tool for detecting disease.
Dynamic optical coherence elastography (OCE) techniques have rapidly emerged as a noninvasive way to characterize the biomechanical properties of tissue. However, clinical applications of the majority of these techniques have been unfeasible due to the extended acquisition time because of multiple temporal OCT acquisitions (M-B mode). Moreover, multiple excitations, large datasets, and prolonged laser exposure prohibit their translation to the clinic, where patient discomfort and safety are critical criteria. Here, we demonstrate the feasibility of noncontact true kilohertz frame-rate dynamic optical coherence elastography by directly imaging a focused air-pulse induced elastic wave with a home-built phase-sensitive OCE system. The OCE system was based on a 4X buffered Fourier Domain Mode Locked swept source laser with an A-scan rate of ~1.5 MHz, and imaged the elastic wave propagation at a frame rate of ~7.3 kHz. Because the elastic wave directly imaged, only a single excitation was utilized for one line scan measurement. Rather than acquiring multiple temporal scans at successive spatial locations as with previous techniques, here, successive B-scans were acquired over the measurement region (B-M mode). Preliminary measurements were taken on tissue-mimicking agar phantoms of various concentrations, and the results showed good agreement with uniaxial mechanical compression testing. Then, the elasticity of an in situ porcine cornea in the whole eye-globe configuration at various intraocular pressures was measured. The results showed that this technique can acquire a depth-resolved elastogram in milliseconds. Furthermore, the ultra-fast acquisition ensured that the laser safety exposure limit for the cornea was not exceeded.
In this study we have evaluated the elastic anisotropy of porcine corneas with increasing intraocular pressures (IOPs) using a noncontact optical coherence elastography (OCE) technique. A focused air-pulse induced low amplitude (≤10 μm) elastic waves in fresh porcine corneas (n=9) in situ in the whole eye-globe configuration. A phase-stabilized swept source optical coherence elastography (PhS-SSOCE) system imaged the propagation of the elastic wave in different stepped radial directions. A closed-loop feedback system was utilized to artificially manipulate the IOP, and OCE measurements were repeated while the IOP was increased in 5 mmHg increments from 15 to 30 mmHg. The OCE measurements demonstrated that the elastic anisotropy of the cornea became more pronounced at higher IOPs, and that there were distinct radial angles of higher and lower stiffness. The presented noncontact OCE method was capable of detecting and assessing the corneal elastic anisotropy as a function of IOP. Due to the noninvasive nature and small amplitude of the elastic wave, this method may be able to provide additional information about corneal health and integrity in vivo.
Dynamic optical coherence elastography (OCE) techniques have shown great promise at quantitatively obtaining the biomechanical properties of tissue. However, the majority of these techniques have required multiple temporal OCT acquisitions (M-B mode) and corresponding excitations, which lead to clinically unfeasible acquisition times and potential tissue damage. Furthermore, the large data sets and extended laser exposures hinder their translation to the clinic, where patient discomfort and safety are critical criteria. In this work we demonstrate noncontact true kilohertz frame-rate dynamic optical coherence elastography by directly imaging a focused air-pulse induced elastic wave with a home-built phase-sensitive OCE system based on a 4X buffered Fourier Domain Mode Locked swept source laser with an A-scan rate of ~1.5 MHz. The elastic wave was imaged at a frame rate of ~7.3 kHz using only a single excitation. In contrast to previous techniques, successive B-scans were acquired over the measurement region (B-M mode) in this work. The feasibility of this method was validated by quantifying the elasticity of tissue-mimicking agar phantoms as well as porcine corneas ex vivo at different intraocular pressures. The results demonstrate that this method can acquire a depth-resolved elastogram in milliseconds. The reduced data set enabled a rapid elasticity assessment, and the ultra-fast acquisition speed allowed for a clinically safe laser exposure to the cornea.
The mechanical anisotropic properties of the cornea can be an important indicator for determining the onset and severity
of different diseases and can be used to assess the efficacy of various therapeutic interventions, such as cross-linking and
LASIK surgery. In this work, we introduce a noncontact method of assessing corneal mechanical anisotropy as a
function of intraocular pressure (IOP) using optical coherence elastography (OCE). A focused air-pulse induced low
amplitude (<10 μm) elastic waves in fresh porcine corneas in the whole eye-globe configuration in situ. A phase-stabilized
swept source optical coherence elastography (PhS-SSOCE) system imaged the elastic wave propagation at
stepped radial angles, and the OCE measurements were repeated as the IOP was cycled. The elastic wave velocity was
then quantified to determine the mechanical anisotropy and hysteresis of the cornea. The results show that the elastic
anisotropy at the corneal of the apex of the cornea becomes more pronounced at higher IOPs, and that there are distinct
radial angles of higher and lower stiffness. Due to the noncontact nature and small amplitude of the elastic wave, this
method may be useful for characterizing the elastic anisotropy of ocular and other tissues in vivo completely
noninvasively.
Elastic wave imaging optical coherence elastography (EWI-OCE) is an emerging technique that can quantify local
biomechanical properties of tissues. However, long acquisition times make this technique unfeasible for clinical use.
Here, we demonstrate a noncontact single shot line-field OCE technique using a line-field interferometer and an air-pulse
delivery system. The spatial-temporal elastic wave propagation profile was acquired in a single shot and used
to quantify the elastic wave group velocity in tissue. Results on tissue-mimicking phantoms and chicken breast
muscle agreed well with mechanical compression testing, demonstrating that the presented method can effectively
reduce the OCE acquisition time to a few milliseconds in biological application.
The murine model is a common model for studying developmental diseases. In this study, we compare the performance of the relatively new method of Optical Projection Tomography (OPT) to the well-established technique of Optical Coherence Tomography (OCT) to assess murine embryonic development at three stages, 9.5, 11.5, and 13.5 days post conception. While both methods can provide spatial resolution at the micrometer scale, OPT can provide superior imaging depth compared to OCT. However, OPT requires samples to be fixed, placed in an immobilization media such as agar, and cleared before imaging. Because OCT does not require fixing, it can be used to image embryos in vivo and in utero. In this study, we compare the efficacy of OPT and OCT for imaging murine embryonic development. The data demonstrate the superior capability of OPT for imaging fine structures with high resolution in optically-cleared embryos while only OCT can provide structural and functional imaging of live embryos ex vivo and in utero with micrometer scale resolution.
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